Inhibitors of cognitive decline

ABSTRACT

Compounds that are central nervous system drug candidates for the treatment of cognitive decline and, more particularly, Alzheimer&#39;s disease are provided. Methods of treating, inhibiting, and/or abatement of cognitive decline and/or Alzheimer&#39;s disease with a compound or pharmaceutically acceptable salt of the invention are also provided. Also provided are methods of preparing the compounds/compositions of the invention.

This application claims benefit of priority to U.S. provisional patentapplication Ser. No. 61/167,984 filed on Apr. 9, 2009, U.S. provisionalpatent application Ser. No. 61/308,667 filed on Feb. 26, 2010, and U.S.provisional patent application Ser. No. 61/309,091 filed on Mar. 1,2010, each of which is hereby incorporated by reference in its entirety.

SUMMARY

The present invention provides, inter alia, compounds of Formula I, II,or III:

or pharmaceutically acceptable salts, wherein constituent members areprovided below.

The present invention further provides pharmaceutical compositionscomprising a compound of Formula I, II, or III, or pharmaceuticallyacceptable salt thereof, and at least one pharmaceutically acceptablecarrier.

The present invention further provides methods of inhibiting, treating,and/or abating cognitive decline and/or Alzheimer's disease with acompound of Formula I, II, or III, or pharmaceutically acceptable saltof the same.

The present invention further provides methods of inhibiting, treating,or abatement of cognitive decline with a compound of Formula I, II, orIII, or pharmaceutically acceptable salt of the same.

The present invention further provides methods of inhibiting, treating,or abatement of one or more of amyloid production, amyloid assembly,amyloid aggregation, amyloid binding (to cells in the brain such asneuron cells), the activity/effect of Abeta oligomers on neurons, andamyloid deposition (on cells in the brain such as neuron cells) with acompound of Formula I, II, or III, or pharmaceutically acceptable saltof the same.

The present invention further provides compounds of Formula I, II, orIII, or pharmaceutically acceptable salts thereof, for use in therapy.

The present invention further provides use of the compounds of FormulaI, II, or III, or pharmaceutically acceptable salts thereof, for themanufacture/preparation of a medicament for use in therapy.

In some embodiments, methods for preparation of compounds useful forinhibiting, treating, or abatement of cognitive decline are provided. Ina method called “chemical conditioning”, certain compounds of thepresent invention are derived from naturally occurring compounds, suchas those found in medicinal plants, like ginger. The chemicalconditioning process described herein is applicable to a large varietyof biological extracts and may be used to create compound arrays forscreening for potential new drug candidates. Further, in general,compounds derived by the chemical conditioning process are chemicallystable and structurally diverse, and good candidates for use in drugscreenings for pharmaceutical activity. In some embodiments, compoundsderived from ginger oil are provided. According to some embodiments ofthe invention, compounds derived from ginger oil by the chemicalconditioning process described herein are provided. In anotherembodiment, the invention provides a method of preparing an array ofchemical compounds from ginger oil.

In some embodiments, the compounds of present invention inhibit, treat,or abate (partially inhibit) binding of the amyloid (including Abetaoligomers) to neurons (such as neurons in the brain) and are useful forthe inhibition, treatment, and abatement of cognitive decline and/orAlzheimer's disease. In some embodiments, the compounds of presentinvention inhibit, treat, or abate one or more of amyloid aggregation,amyloid binding, and amyloid deposition. In some embodiments, thecompounds of present invention inhibit, treat, or abate amyloidaggregation. In some embodiments, the compounds of present inventioninhibit, treat, or abate amyloid binding. In some embodiments, thecompounds of present invention inhibit, treat, or abate amyloiddeposition. In some embodiments, the compounds of present inventioninhibit, treat, or abate the activity/effect of Abeta oligomers onneurons. In some embodiments, the compounds show activity in abeta-secretase assay and are potentially useful for the inhibition,treatment, and abatement of cognitive decline and Alzheimer's disease.In some embodiments the derivative of ginger oil is a compound inpurified and isolated form (for example, with a purity of greater than80%, 85%, 90%, 95%, 98%, or 99% by weight). The compounds and methodsdescribed herein may be used to treat one or more symptoms of cognitivedecline and/or Alzheimer's disease such as memory loss, confusion,impaired judgment, personality changes, disorientation, and loss oflanguage skills. Further, the compounds and methods described herein maybe useful in inhibiting, treating, and/or abating cognitive declineand/or Alzheimer's disease by restoring long term potentiation, and/orinhibiting, treating, or abatement of one or both of neurodegenerationand general amyloidosis, more specifically, by inhibiting, treating, orabatement of one or more of amyloid production, amyloid assembly,amyloid aggregation, amyloid binding, and amyloid deposition.

DESCRIPTION OF DRAWINGS

FIG. 1 shows results of an MTT assay in the presence and absence of aprocessed product of amyloid precursor protein.

FIG. 2 shows inhibition of processed product of amyloid precursorprotein-mediated membrane trafficking effect by Compound Example 2.

FIG. 3 shows Compound Example 2 inhibiting the memory loss effects of aprocessed product of amyloid precursor protein.

FIG. 4 shows Compound Example 2 Inhibiting the membrane traffickingeffects of Abeta assemblies isolated from AD patients.

DETAILED DESCRIPTION

Cognitive decline, such as memory loss, confusion, impaired judgment,personality changes, disorientation, and loss of language skills occursin much of the population as they age, in varying degree. The mostcommon, severe and irreversible form of cognitive decline is Alzheimer'sdisease, which, at present, is always fatal.

The symptoms of cognitive decline and Alzheimer's disease are thought tostem from the formation of amyloid plaques and neurofibrillary tangles,which are thought to contribute to the degradation of the neurons (nervecells) in the brain and the subsequent onset of symptoms. Amyloid is ageneral term for protein fragments that the body produces normally.Beta-amyloid is a fragment of a protein that is snipped from anotherprotein called amyloid precursor protein (APP). In a healthy brain,beta-amyloid protein fragments are broken down and eliminated. Inindividuals with Alzheimer's disease and other forms of cognitivedecline, the fragments accumulate to form hard, insoluble plaques.Neurofibrillary tangles are insoluble twisted fibers that are foundinside of the brain's cells. The protein contained in neurofibrillarytangles, i.e., the tau protein, forms a microtubule, which helpstransport nutrients and other important substances from one part of thenerve cell to another. In Alzheimer's disease the tau protein isabnormal and the microtubule structures collapse.

Beta-secretase is the enzyme in the human brain responsible for theproduction of Beta-amyloid, the pathogenic substance responsible for theformation of brain plaques and tangles in the Alzheimer's diseasedbrain. Beta-amyloid and its oligomers (beta-amyloid oligomers or Abetaoligomers) are also believed to be responsible for early cognitivedecline in the pre-Alzheimer's diseased brain. Inhibition ofbeta-secretase would be expected to lessen beta-amyloid burden in thebrain and thus slow cognitive decline, block the formation of amyloidoligomers, the production of plaques and tangles, haltneurodegeneration, and to potentially treat mild cognitive impairmentand more serious forms of cognitive impairment such as Alzheimer'sdisease.

The gingerols are a series of natural small molecules isolated fromginger, Zingiber officinale, and are classified according to their alkylchain length e.g., [6]-gingerol, [8]-gingerol. Gingerols are known to berelatively unstable under both chemical and biological conditions,forming Inactive substances. For example, the beta-hydroxycarbonylfunction of the gingerols is vulnerable to oxidation or dehydration toform inactive products, and the gingerols are particularly prone torapid dehydration under acidic conditions, such that even the puresubstance is difficult to store for long periods. Accordingly, simpleoral dosing of the gingerols for medicinal action might not be possibledue to the acidic environment of the stomach and upper intestinal tract.Further, chemical and biological instability is also likely to be aserious problem for intravenous doses. Accordingly, there is strong needto discover inhibitors of cognitive decline, and in particular,compounds that are useful in the treatment and abatement of cognitivedecline and Alzheimer's disease, by methods such as inhibiting amyloid(including Abeta oligomers) production, amyloid (including Abetaoligomers) aggregation, and/or amyloid (including Abets oligomers)deposition (i.e., plaqing), inhibiting neuorodegeneration, and/orrestoring long term potentiation, and/or Inhibiting the activity/effectof Abeta oligomers on neurons. There is also a need for inhibitors ofcognitive decline that are chemically and biologically stable.

Plants have attracted relatively little attention as potentiallyvaluable resources for drug discovery in the area of cognitive declineand Alzheimer's disease. The use of plant extracts to produce unnaturalderivatives of compounds of medicinal interest is not generally used.Accordingly, there is also a need for a method of producing compounds ofmedicinal interest from plant extracts and extracts from otherbiological sources. In particular, there is also a need to produce andidentify compounds derived from plant extracts that are useful in thetreatment and abatement of cognitive decline and Alzheimer's disease.

The compounds, compositions, and methods described herein are directedtoward these needs and other ends.

Embodiments of the present invention provides, inter alia, compounds ofFormula I:

or pharmaceutically acceptable salts thereof wherein:

R¹ is selected from (A1) and (A2):

R², R³, R⁴, R⁵, and R⁶ are each, independently, selected from H, OH,C₁₋₆ alkoxy, C₁₋₆ haloalkoxy, halo, CN, NO₂, C₁₋₆ alkyl, C₁₋₆ haloalkyl,C₃₋₇cycloalkyl, NH₂, NH(C₁₋₄ alkyl), NH(C₃₋₇ cycloalkyl), N(C₁₋₄alkyl)₂, NHC(O)(C₁₋₄ alkyl), SH, S(C₁₋₆ alkyl), C(O)OR^(a), C(O)R^(b),C(O)NR^(c)R^(d), OC(O)R^(b), OC(O)NR^(c)R^(d), NR^(c)R^(d),NR^(c)C(O)R^(b), NR^(c)C(O)OR^(a), NR^(c)S(O)₂R^(b),NR^(c)S(O)₂NR^(c)R^(d), S(O)R^(b), S(O)₂R^(b), and S(O)₂NR^(c)R^(d);

R⁷ is H, C₁₋₆ alkyl, C₁₋₆ haloalkyl, or C₃₋₇ cycloalkyl;

R⁸ is C₁₋₆ alkyl, C₁₋₆ haloalkyl, or C₃₋₇ cycloalkyl;

R⁹ is H, C₁₋₆ alkyl, C₁₋₆ haloalkyl, or C₃₋₇ cycloalkyl;

R¹⁰ is H, C₁₋₆ alkyl, C₁₋₆ haloalkyl, or C₃₋₇ cycloalkyl;

R¹¹ is H, C₁₋₆ alkyl, C₁₋₆ haloalkyl, or C₃₋₇ cycloalkyl;

R¹², R¹³, R¹⁴, R¹⁵, and R¹⁶ are each, independently, selected from H,OH, C₁₋₆ alkoxy, C₁₋₆ haloalkoxy, halo, CN, NO₂, C₁₋₆ alkyl, C₁₋₆haloalkyl, C₃₋₇ cycloalkyl, NH₂, NH(C₁₋₄ alkyl), NH(C₃₋₇ cycloalkyl),N(C₁₋₄ alkyl)₂, NHC(O)(C₁₋₄ alkyl), SH, S(C₁₋₆ alkyl), C(O)OR^(a1),C(O)R^(b1), C(O)NR^(c1)R^(d1), OC(O)R^(b1), OC(O)NR^(c1)R^(d1),NR^(e1)R^(d1), NR^(c1)C(O)R^(b1), NR^(c1)C(O)OR^(a1),NR^(c1)S(O)₂R^(b1), NR^(c1)S(O)₂NR^(c1)R^(d1), S(O)R^(b1), S(O)R^(b1),and S(O)₂NR^(c1)R^(d1);

each R^(a) is independently selected from H, C₁₋₆ alkyl, C₁₋₆ haloalkyl,arylalkyl, heteroarylalkyl, cycloalkylalkyl, heterocycloalkylalkyl, C₂₋₆alkenyl, C₂₋₆ alkynyl, aryl, cycloalkyl, heteroaryl andheterocycloalkyl, wherein each of the C₁₋₆ alkyl, C₁₋₆ haloalkyl,arylalkyl, heteroarylalkyl, cycloalkylalkyl, heterocycloalkylalkyl, C₂₋₆alkenyl, C₂₋₆ alkynyl, aryl, cycloalkyl, heteroaryl and heterocycloalkylis optionally substituted with 1, 2, 3, 4, or 5 substituentsindependently selected from OH, CN, amino, halo, C₁₋₆ alkyl, C₁₋₆haloalkyl, C₁₋₆ alkoxy, C₁₋₆ haloalkoxy, aryl, arylalkyl, heteroaryl,heteroarylalkyl, cycloalkyl and heterocycloalkyl;

each R^(b) is independently selected from H, C₁₋₆ alkyl, C₁₋₆ haloalkyl,arylalkyl, heteroarylalkyl, cycloalkylalkyl, heterocycloalkylalkyl, C₂₋₆alkenyl, C₂₋₆ alkynyl, aryl, cycloalkyl, heteroaryl, heterocycloalkyl,arylalkyl, heteroarylalkyl, cycloalkylalkyl and heterocycloalkylalkyl,wherein each of the C₁₋₆ alkyl, C₁₋₆ haloalkyl, arylalkyl,heteroarylalkyl, cycloalkylalkyl, heterocycloalkylalkyl, C₂₋₆ alkenyl,C₂₋₆ alkynyl, aryl, cycloalkyl, heteroaryl, heterocycloalkyl, arylalkyl,heteroarylalkyl, cycloalkylalkyl and heterocycloalkylalkyl is optionallysubstituted with 1, 2, 3, 4, or 5 substituents independently selectedfrom OH, amino, halo, C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₁₋₆ alkoxy, C₁₋₆haloalkoxy, aryl, arylalkyl, heteroaryl, heteroarylalkyl, cycloalkyl andheterocycloalkyl;

R^(c) and R^(d) are independently selected from H, C₁₋₆ alkyl, C₁₋₆haloalkyl, arylalkyl, heteroarylalkyl, cycloalkylalkyl,heterocycloalkylalkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl, aryl, heteroaryl,cycloalkyl, heterocycloalkyl, arylalkyl, heteroarylalkyl,cycloalkylalkyl and heterocycloalkylalkyl, wherein each of the C₁₋₆alkyl, C₁₋₆ haloalkyl, arylalkyl, heteroarylalkyl, cycloalkylalkyl,heterocycloalkylalkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl, aryl, heteroaryl,cycloalkyl, heterocycloalkyl, arylalkyl, heteroarylalkyl,cycloalkylalkyl and heterocycloalkylalkyl is optionally substituted with1, 2, 3, 4, or 5 substituents independently selected from OH, amino,halo, C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₁₋₆ alkoxy, C₁₋₆ haloalkoxy, aryl,arylalkyl, heteroaryl, heteroarylalkyl, cycloalkyl and heterocycloalkyl;

or R^(c) and R^(d) together with the N atom to which they are attachedform a 4-, 5-, 6- or 7-membered heterocycloalkyl group that isoptionally substituted with 1, 2, 3, 4, or 5 substituents independentlyselected from OH, amino, halo, C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₁₋₆ alkoxy,C₁₋₆ haloalkoxy, aryl, arylalkyl, heteroaryl, heteroarylalkyl,cycloalkyl, and heterocycloalkyl;

-   -   each R^(a1) is independently selected from H, C₁₋₆ alkyl, C₁₋₆        haloalkyl, arylalkyl, heteroarylalkyl, cycloalkylalkyl,        heterocycloalkylalkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl, aryl,        cycloalkyl, heteroaryl and heterocycloalkyl, wherein each of the        C₁₋₆ alkyl, C₁₋₆ haloalkyl, arylalkyl, heteroarylalkyl,        cycloalkylalkyl, heterocycloalkylalkyl, C₂₋₆ alkenyl, C₂₋₆        alkynyl, aryl, cycloalkyl, heteroaryl and heterocycloalkyl is        optionally substituted with 1, 2, 3, 4, or 5 substituents        independently selected from OH, CN, amino, halo, C₁₋₆ alkyl,        C₁₋₆ haloalkyl, C₁₋₆ alkoxy, C₁₋₆ haloalkoxy, aryl, arylalkyl,        heteroaryl, heteroarylalkyl, cycloalkyl, and heterocycloalkyl;

each R^(b1) is independently selected from H, C₁₋₆ alkyl, C₁₋₆haloalkyl, arylalkyl, heteroarylalkyl, cycloalkylalkyl,heterocycloalkylalkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl, aryl, cycloalkyl,heteroaryl, heterocycloalkyl, arylalkyl, heteroarylalkyl,cycloalkylalkyl and heterocycloalkylalkyl, wherein each of the C₁₋₆alkyl, C₁₋₆ haloalkyl, arylalkyl, heteroarylalkyl, cycloalkylalkyl,heterocycloalkylalkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl, aryl, cycloalkyl,heteroaryl, heterocycloalkyl, arylalkyl, heteroarylalkyl,cycloalkylalkyl and heterocycloalkylalkyl is optionally substituted with1, 2, 3, 4, or 5 substituents independently selected from OH, amino,halo, C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₁₋₆ alkoxy, C₁₋₆ haloalkoxy, aryl,arylalkyl, heteroaryl, heteroarylalkyl, cycloalkyl, andheterocycloalkyl;

R^(c1) and R^(d2) are independently selected from H, C₁₋₆ alkyl, C₁₋₆haloalkyl, arylalkyl, heteroarylalkyl, cycloalkylalkyl,heterocycloalkylalkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl, aryl, heteroaryl,cycloalkyl, heterocycloalkyl, arylalkyl, heteroarylalkyl,cycloalkylalkyl and heterocycloalkylalkyl, wherein each of the C₁₋₆alkyl, C₁₋₆ haloalkyl, arylalkyl, heteroarylalkyl, cycloalkylalkyl,heterocycloalkylalkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl, aryl, heteroaryl,cycloalkyl, heterocycloalkyl, arylalkyl, heteroarylalkyl,cycloalkylalkyl and heterocycloalkylalkyl is optionally substituted with1, 2, 3, 4, or 5 substituents Independently selected from OH, amino,halo, C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₁₋₆ alkoxy, C₁₋₆ haloalkoxy, aryl,arylalkyl, heteroaryl, heteroarylalkyl, cycloalkyl, andheterocycloalkyl;

-   -   or R^(c1) and R^(d1) together with the N atom to which they are        attached form a 4-, 5-, 6- or 7-membered heterocycloalkyl group        that is optionally substituted with 1, 2, 3, 4, or 5        substituents independently selected from OH, amino, halo, C₁₋₆        alkyl, C₁₋₆ haloalkyl, C₁₋₆ alkoxy, C₁₋₆ haloalkoxy, aryl,        arylalkyl, heteroaryl, heteroarylalkyl, cycloalkyl, and        heterocycloalkyl; and

m is 0, 1, or 2.

In some embodiments, when R¹ is a moiety of (A1), then two of R², R³,R⁴, R⁵, and R⁶ are independently selected from OH, C₁₋₆ alkoxy, and C₁₋₆haloalkoxy.

In some embodiments, when R¹ is a moiety of (A1), then at least one ofR¹², R¹³, R¹⁴, R¹⁵, and R¹⁶ is other than H.

In some embodiments, two of R², R³, R⁴, R⁵, and R⁶ are independentlyselected from OH, C₁₋₆ alkoxy, and C₁₋₆ haloalkoxy. In some furtherembodiments, each of the rest of R², R³, R⁴, R⁵, and R⁶ is H.

In some embodiments, R², R³, R⁴, R⁵, and R⁶ are each, independently,selected from H, OH, C₁₋₆ alkoxy, C₁₋₆ haloalkoxy, halo, CN, NO₂, C₁₋₆alkyl, C₁₋₆ haloalkyl, C₃₋₇ cycloalkyl, NH₂, NH(C₁₋₄ alkyl), NH(C₃₋₇cycloalkyl), N(C₁₋₄ alkyl)₂, NHC(O)(C₁₋₄ alkyl), SH, S(C₁₋₆ alkyl),C(O)OH, C(O)O(C₁₋₄ alkyl), C(O)(C₁₋₄ alkyl), and C(O)NH(C₁₋₄ alkyl).

In some embodiments, one of R², R³, R⁴, R⁵, and R⁶ is OH; and one of R²,R³, R⁴, R⁵, and R⁶ is OH, C₁₋₆ alkoxy, or C₁₋₆ haloalkoxy. In somefurther embodiments, each of the rest of R², R³, R⁴, R⁵, and R⁶ is H.

In some embodiments, one of R², R³, R⁴, R⁵, and R⁶ is OH; and one of R²,R³, R⁴, R⁵, and R⁶ is C₁₋₃ alkoxy or C₁₋₃ haloalkoxy (In some furtherembodiments, each of the rest of R², R³, R⁴, R⁵, and R⁶ is H.). In somefurther embodiments, one of R², R³, R⁴, R⁵ and R⁶ is OH; and one of R²,R³, R⁴, R⁵, and R⁶ is methoxy or trihalomethoxy (In some furtherembodiments, each of the rest of R², R³, R⁴, R⁵, and R⁶ is H.). In stillfurther embodiments, one of R², R³, R⁴, R⁵, and R⁶ is OH; and one of R²,R³, R⁴, R⁵, and R⁶ is methoxy (In some further embodiments, each of therest of R², R³, R⁴, R⁵, and R⁶ is H.).

In some embodiments, R⁴ is OH; and R⁵ is methoxy. In some furtherembodiments, R⁴ is OH; R⁵ is methoxy; and R², R³, and R⁶ are each H.

In some embodiments, R⁷ is H or C₁₋₆ alkyl. In some further embodiments,R⁷ is H or C₁₋₃ alkyl.

In some embodiments, R⁷ is C₁₋₃ alkyl. In some further embodiments, R⁷is methyl or ethyl. In still further embodiments, R⁷ is methyl.

In some embodiments, R⁷ is H.

In some embodiments, R⁸ is C₁₋₆ alkyl. In some further embodiments, R⁸is C₁₋₃ alkyl. In still further embodiments, R⁸ is methyl.

In some embodiments, R⁹ is H or C₁₋₆ alkyl. In some further embodiments,R⁹ is H or C₁₋₃ alkyl.

In some embodiments, R⁹ is H.

In some embodiments, R⁹ is C₁₋₃ alkyl.

In some embodiments, R¹⁰ is H or C₁₋₆ alkyl. In some furtherembodiments, R¹⁰ is H or C₁₋₃ alkyl. In still further embodiments, R¹⁰is H. In other embodiments, R¹⁰ is C₁₋₃ alkyl.

In some embodiments, R¹¹ is H or C₁₋₆ alkyl. In some furtherembodiments, R¹¹ is H or C₁₋₃ alkyl. In still further embodiments. R¹¹is H. In other embodiments, R¹¹ is C₁₋₃ alkyl.

In some embodiments, at least one of R¹², R¹³, R¹⁴, R¹⁵, and R¹⁶ isother than H.

In some embodiments, R¹², R¹³, R¹⁴, R¹⁵, and R¹⁶ are each,independently, selected from H, OH, C₁₋₆ alkoxy, C₁₋₆ haloalkoxy, halo,CN, NO₂, C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₃₋₇ cycloalkyl, NH₂, NH(C₁₋₄alkyl), NH(C₃₋₇cycloalkyl), N(C₁₋₄ alkyl)₂, NHC(O)(C₁₋₄ alkyl), SH,S(C₁₋₆ alkyl), C(O)OH, C(O)O(C₁₋₄ alkyl), C(O)(C₁₋₄ alkyl), andC(O)NH(C₁₋₄ alkyl).

In some embodiments, R¹², R¹³, R¹⁴, R¹⁵, and R¹⁶ are each,independently, selected from H, halo, CN, NO₂, C₁₋₆ alkyl, C₁₋₆haloalkyl, C₃₋₇ cycloalkyl, C(O)O(C₁₋₄ alkyl), C(O)(C₁₋₄ alkyl), andC(O)NH(C₁₋₄ alkyl).

In some embodiments, at least one of R¹², R¹³, R¹⁴, R¹⁵, and R¹⁶ isselected from halo, CN, NO₂, C₁₋₆ haloalkyl, C(O)O(C₁₋₄ alkyl),C(O)(C₁₋₄ alkyl), and C(O)NH(C₁₋₄ alkyl).

In some embodiments, at least one of R¹², R¹³, R¹⁴, R¹⁵, and R¹⁶ isselected from halo and C₁₋₆ haloalkyl. In some further embodiments, atleast one of R¹², R¹³, R¹⁴, R¹⁵, and R¹⁶ is selected from halo and C₁₋₆haloalkyl, and each of the rest is of R¹², R¹³, R¹⁴, R¹⁵, and R¹⁶ is H.In yet further embodiments, one or two of R¹², R¹³, R¹⁴, R¹⁵, and R¹⁶are selected from halo and C₁₋₆ haloalkyl, and each of the rest is ofR¹², R¹³, R¹⁴, R¹⁵, and R¹⁶ is H. In still further embodiments, one ofR¹², R¹³, R¹⁴, R¹⁵, and R¹⁸ is selected from halo and C₁₋₆ haloalkyl,and each of the rest is of R¹², R¹³, R¹⁴, R¹⁵, and R¹⁶ is H.

In some embodiments, R¹⁴ is halo or C₁₋₆ haloalkyl (In some furtherembodiments, each of R¹², R¹³, R¹⁵, and R¹⁶ is H.). In some furtherembodiments, R¹⁴ is halo or C₁₋₃ haloalkyl (In some further embodiments,each of R¹², R¹³, R¹⁵, and R¹⁶ is H.). In still further embodiments, R¹⁴is halo or C₁ haloalkyl (In some further embodiments, each of R¹², R¹³,R¹⁵, and R¹⁶ is H.).

In some embodiments, R¹⁴ is halo (In some further embodiments, each ofR¹², R¹³, R¹⁵, and R¹⁶ is H.). In some embodiments, R¹⁴ is Cl or F. Insome embodiments, R¹⁴ is Cl. In some embodiments, R¹⁴ is F.

In some embodiments, R¹⁴ is C₁₋₆ haloalkyl (In some further embodiments,each of R¹², R¹³, R¹⁵, and R¹⁶ is H.). In some further embodiments, R¹⁴is C₁₋₃ haloalkyl. In still further embodiments, R¹⁴ is C₁ haloalkyl. Inyet further embodiments, R¹⁴ is CF₃.

In some embodiments, R¹⁵ is halo or C₁₋₆ haloalkyl (In some furtherembodiments, each of R¹², R¹³, R¹⁴, and R¹⁶ is H.). In some furtherembodiments, R¹⁵ is halo or C₁₋₃ haloalkyl (In some further embodiments,each of R¹², R¹³, R¹⁴, and R¹⁶ is H.). In still further embodiments, R¹⁵is halo or C₁ haloalkyl (In some further embodiments, each of R¹², R¹³,R¹⁴, and R¹⁶ is H.).

In some embodiments, R¹⁵ is halo. In some embodiments, R¹⁵ is Cl or F.In some embodiments, R¹⁵ is Cl. In some embodiments, R¹⁵ is F.

In some embodiments, R¹⁴ is C₁₋₆ haloalkyl. In some further embodiments,R¹⁵ is C₁₋₃ haloalkyl. In still further embodiments, R¹⁵ is C₁haloalkyl. In yet further embodiments, R¹⁵ is CF₃.

In some embodiments, R¹⁴ and R¹⁵ are each independently halo or C₁₋₃haloalkyl (In some further embodiments, each of R¹², R¹³, and R¹⁶ isH.). In some further embodiments, R¹⁴ and R¹⁵ are each independentlyhalo or C₁ haloalkyl.

In some embodiments, R¹⁴ and R¹⁵ are each independently halo.

In some embodiments, the compound of Formula I is a compound of FormulaII:

In some embodiments, the compound of Formula II or pharmaceuticallyacceptable suit thereof is a compound of Formula IIa or IIb:

or pharmaceutically acceptable salt thereof.

In some embodiments, the compound of Formula II is a compound of FormulaIIa. In some further embodiments, R¹⁰ and R¹¹ are each, independently,selected from H and C₁₋₃ alkyl. In yet further embodiments, R¹⁰ and R¹¹are each, independently, selected from H and methyl. In still furtherembodiments, R¹⁰ and R¹¹ are each H.

In some embodiments of the compound of Formula IIa or pharmaceuticallyacceptable salt thereof, one of R¹⁰ and R¹¹ is selected from H and C₁₋₃alkyl and the other is H. In some further embodiments, one of R¹⁰ andR¹¹ is C₁₋₃ alkyl. In yet further embodiments, one of R¹⁰ and R¹¹ ismethyl.

In some embodiments of the compound of Formula IIa or pharmaceuticallyacceptable salt thereof, both of R¹⁰ and R¹¹ are selected from C₁₋₃alkyl. In some further embodiments, both R¹⁰ and R¹¹ are methyl.

In some embodiments of the compound of Formula IIa or pharmaceuticallyacceptable salt thereof, R¹², R¹³, R¹⁴, R¹⁵, and R¹⁶ are each,independently, selected from H, OH, C₁₋₆ alkoxy, C₁₋₆ haloalkoxy, halo,CN, NO₂, C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₃₋₇ cycloalkyl, NH₂, NH(C₁₋₄alkyl), NH(C₃₋₇cycloalkyl), N(C₁₋₄ alkyl)₂, NHC(O)(C₁₋₄ alkyl), SH,S(C₁₋₆ alkyl), C(O)OH, C(O)O(C₁₋₄ alkyl), C(O)(C₁₋₄ alkyl), andC(O)NH(C₁₋₄ alkyl).

In some embodiments of the compound of Formula IIa or pharmaceuticallyacceptable salt thereof, R¹², R¹³, R¹⁴, R¹⁵, and R¹⁶ are each,independently, selected from H, halo, CN, NO₂, C₁₋₆ alkyl, C₁₋₆haloalkyl, C₃₋₇ cycloalkyl, C(O)O(C₁₋₄ alkyl), C(O)(C₁₋₄ alkyl), andC(O)NH(C₁₋₄ alkyl).

In some embodiments of the compound of Formula IIa or pharmaceuticallyacceptable salt thereof, R¹², R¹³, R¹⁴, R¹⁵, and R¹⁶ are each,independently, selected from H, halo, CN, NO₂, C₁₋₆ alkyl, C₁₋₆haloalkyl, and C₃₋₇ cycloalkyl. In some further embodiments, R¹², R¹³,R¹⁴, R¹⁵, and R¹⁶ are each, independently, selected from H, halo, CN,C₁₋₆ alkyl, and C₁₋₆ haloalkyl. In yet further embodiments, R¹², R¹³,R¹⁴, R¹⁵, and R¹⁶ are each, independently, selected from H, halo, C₁₋₆alkyl, and C₁₋₆ haloalkyl.

In some embodiments of the compound of Formula IIa or pharmaceuticallyacceptable salt thereof, at least one of R¹², R¹³, R¹⁴, R¹⁵, and R¹⁶ isselected from halo and C₁₋₆ haloalkyl, and each of the rest is of R¹²,R¹³, R¹⁴, R¹⁵, and R¹⁶ is H. In some further embodiments, one or two ofR¹², R¹³, R¹⁴, R¹⁵, and R¹⁶ are selected from halo and C₁₋₆ haloalkyl,and each of the rest is of R¹², R¹³, R¹⁴, R¹⁵, and R¹⁶ is H. In yetfurther embodiments, one of R¹², R¹³, R¹⁴, R¹⁵, and R¹⁶ is selected fromhalo and C₁₋₆ haloalkyl, and each of the rest is of R¹², R¹³, R¹⁴, R¹⁵,and R¹⁶ is H.

In some embodiments of the compound of Formula IIa or pharmaceuticallyacceptable salt thereof, at least one of R¹², R¹³, R¹⁴, R¹³, and R¹¹ isselected from halo, CN, NO₂, C₁₋₆ haloalkyl, C(O)O(C₁₋₄ alkyl),C(O)(C₁₋₄ alkyl), and C(O)NH(C₁₋₄ alkyl).

In some embodiments of the compound of Formula IIa, at least one of R¹²,R¹³, R¹⁴, R¹⁵, and R¹⁶ is selected from halo and C₁₋₆ haloalkyl.

In some embodiments of the compound of Formula IIa or pharmaceuticallyacceptable salt thereof, at least one of R¹², R¹³, R¹⁴, R¹⁵, and R¹⁶ isselected from halo and C₁₋₃ haloalkyl. In some further embodiments, atleast one of R¹², R¹³, R¹⁴, R¹⁵, and R¹⁶ is selected from halo and C₁haloalkyl.

In some embodiments of the compound of Formula IIa or pharmaceuticallyacceptable salt thereof, R¹⁴ is halo or C₁₋₆ haloalkyl. In some furtherembodiments, R¹⁴ is halo or C₁₋₃ haloalkyl. In still furtherembodiments, R¹⁴ is halo or C₁ haloalkyl.

In some embodiments of the compound of Formula IIa or pharmaceuticallyacceptable salt thereof, R¹⁴ is halo (In some further embodiments, eachof R¹², R¹³, R¹⁵, and R¹⁶ is H.). In some embodiments, R¹⁴ is Cl or F.In some embodiments, R¹⁴ is Cl. In some embodiments, R¹⁴ is F.

In some embodiments of the compound of Formula IIa, R¹⁴ is C₁₋₆haloalkyl (In some further embodiments, each of R¹², R¹³, R¹⁵, and R¹⁶is H.). In some further embodiments, R¹⁴ is C₁₋₃ haloalkyl. In stillfurther embodiments, R¹⁴ is C₁ haloalkyl. In yet further embodiments,R¹⁴ is CF₃.

In some embodiments of the compound of Formula IIa or pharmaceuticallyacceptable salt thereof, R¹⁴ is halo or C₁₋₆ haloalkyl and each of R¹²,R¹³, R¹⁵, and R¹⁶ is H.

In some embodiments of the compound of Formula IIa or pharmaceuticallyacceptable salt thereof, R¹⁵ is halo or C₁₋₆ haloalkyl (In some furtherembodiments, each of R¹², R¹³, R¹⁴, and R⁶ is H.). In some furtherembodiments, R¹⁵ is halo or C₁₋₃ haloalkyl. In still furtherembodiments, R¹⁵ is halo or C₁ haloalkyl.

In some embodiments of the compound of Formula IIa or pharmaceuticallyacceptable salt thereof, R¹⁵ is halo. In some embodiments, R¹⁵ is Cl orF. In some embodiments, R¹⁵ is Cl. In some embodiments, R¹¹ is F.

In some embodiments of the compound of Formula IIa or pharmaceuticallyacceptable salt thereof, R¹⁴ is C₁₋₆ haloalkyl. In some furtherembodiments, R¹⁵ is C₁₋₃ haloalkyl. In still further embodiments, R¹⁵ isC₁ haloalkyl. In yet further embodiments, R¹⁵ is CF₃.

In some embodiments of the compound of Formula IIa or pharmaceuticallyacceptable salt thereof, R¹⁴ and R¹⁵ are each independently halo or C₁₋₃haloalkyl (In some further embodiments, each of R¹², R¹³, and R¹⁶ isH.). In some further embodiments, R¹⁴ and R¹⁵ are each independentlyhalo or C₁ haloalkyl. In yet further embodiments, R¹⁴ and R¹⁵ are eachindependently halo.

In some embodiments, the compound of Formula II or pharmaceuticallyacceptable salt thereof is a compound of Formula IIb or pharmaceuticallyacceptable salt thereof.

In some embodiments of the compound of Formula IIb or pharmaceuticallyacceptable salt thereof, R¹², R¹³, R¹⁴, R¹⁵, and R¹⁶ are each,independently, selected from H, OH, C₁₋₆ alkoxy, C₁₋₆ haloalkoxy, halo,CN, NO₂, C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₃₋₇cycloalkyl, NH₂, NH(C₁₋₄alkyl), NH(C₁₋₄ cycloalkyl), N(C₁₋₄ alkyl)₂, NHC(O)(C₁₋₁₄ alkyl), SH,S(C_(1-s) alkyl), C(O)OH, C(O)O(C₁₋₄ alkyl), C(O)(C₁₋₄ alkyl), andC(O)NH(C₁₋₄ alkyl).

In some embodiments of the compound of Formula IIb or pharmaceuticallyacceptable salt thereof, R¹², R¹³, R¹⁴, R¹⁵, and R¹⁶ are each,independently, selected from H, halo, CN, NO₂, C₁₋₆ alkyl, C₁₋₆haloalkyl, C₃₋₇ cycloalkyl, C(O)O(C₁₋₄ alkyl), C(O)(C₁₋₄ alkyl), andC(O)NH(C₁₋₄ alkyl).

In some embodiments of the compound of Formula IIb or pharmaceuticallyacceptable salt thereof, R¹², R¹³, R¹⁴, R¹⁵, and R¹⁶ are each,independently, selected from H, halo, CN, NO₂, C₁₋₆ alkyl, C₁₋₆haloalkyl, and C₃₋₇ cycloalkyl. In some further embodiments, R¹², R¹³,R¹⁴, R¹⁵, and R¹⁶ are each, independently, selected from H, halo, CN,C₁₋₆ alkyl, and C₁₋₆ haloalkyl. In yet further embodiments, R¹², R¹³,R¹⁴, R¹⁵, and R¹⁶ are each, independently, selected from H, halo, C₁₋₆alkyl, and C₁₋₆ haloalkyl.

In some embodiments of the compound of Formula IIb or pharmaceuticallyacceptable salt thereof, at least one of R¹², R¹³, R¹⁴, R¹⁵, and R¹⁶ isselected from halo and C₁₋₆ haloalkyl, and each of the rest is of R¹²,R¹³, R¹⁴, R¹⁵, and R¹⁶ is H. In some further embodiments, one or two ofR¹², R¹³, R¹⁴, R¹⁵, and R¹⁶ are selected from halo and C₁₋₆ haloalkyl,and each of the rest is of R¹², R¹³, R¹⁴, R¹⁵, and R¹⁶ is H. In yetfurther embodiments, one of R¹², R¹³, R¹⁴, R¹⁵, and R¹⁶ is selected fromhalo and C₁₋₆ haloalkyl, and each of the rest is of R¹², R¹³, R¹⁴, R¹⁵,and R¹⁶ is H.

In some embodiments of the compound of Formula IIb or pharmaceuticallyacceptable salt thereof, at least one of R¹², R¹³, R¹⁴, R¹⁵, and R¹⁶ isselected from halo, CN, NO₂, C₁₋₆ haloalkyl, C(O)O(C₁₋₄ alkyl),C(O)(C₁₋₄ alkyl), and C(O)NH(C₁₋₄ alkyl).

In some embodiments of the compound of Formula IIb, at least one of R¹²,R¹³, R¹⁴, R¹⁵, and R¹⁶ is selected from halo and C₁₋₆ haloalkyl.

In some embodiments of the compound of Formula IIb or pharmaceuticallyacceptable salt thereof, at least one of R¹², R¹³, R¹⁴, R¹⁵, and R¹⁶ isselected from halo and C₁₋₃ haloalkyl. In some further embodiments, atleast one of R¹², R¹³, R¹⁴, R¹⁵, and R¹⁶ is selected from halo and C₁haloalkyl.

In some embodiments of the compound of Formula IIb or pharmaceuticallyacceptable salt thereof, R¹⁴ is halo or C₁₋₆ haloalkyl. In some furtherembodiments, R¹⁴ is halo or C₁₋₃ haloalkyl. In still furtherembodiments, R¹⁴ is halo or C₁ haloalkyl.

In some embodiments of the compound of Formula IIb or pharmaceuticallyacceptable salt thereof, R¹⁴ is halo (In some further embodiments, eachof R¹², R¹³, R¹⁵, and R¹⁶ is H.). In some embodiments, R¹⁴ is Cl or F.In some embodiments, R¹⁴ is Cl. In some embodiments, R¹⁴ is F.

In some embodiments of the compound of Formula IIb, R¹⁴ is C₁₋₆haloalkyl (In some further embodiments, each of R¹², R¹³, R¹⁵, and R¹⁶is H.). In some further embodiments, R¹⁴ is C₁₋₃ haloalkyl. In stillfurther embodiments, R¹⁴ is C₁ haloalkyl. In yet further embodiments,R¹⁴ is CF₃.

In some embodiments of the compound of Formula IIb or pharmaceuticallyacceptable salt thereof, R¹⁴ is halo or C₁₋₆ haloalkyl and each of R¹²,R¹³, R¹⁵, and R¹⁶ is H.

In some embodiments of the compound of Formula IIb or pharmaceuticallyacceptable salt thereof, R¹⁵ is halo or C₁₋₆ haloalkyl (In some furtherembodiments, each of R¹², R¹³, R¹⁴, and R¹⁶ is H.). In some furtherembodiments, R¹⁵ is halo or C₁₋₃ haloalkyl. In still furtherembodiments, R¹⁵ is halo or Cl haloalkyl.

In some embodiments of the compound of Formula IIb or pharmaceuticallyacceptable salt thereof, R¹⁵ is halo. In some embodiments, R¹⁵ is Cl orF. In some embodiments, R¹⁵ is Cl. In some embodiments, R¹⁵ is F.

In some embodiments of the compound of Formula IIb or pharmaceuticallyacceptable salt thereof, R¹⁵ is C₁₋₆ haloalkyl. In some furtherembodiments. R¹⁵ is C₁₋₃ haloalkyl. In still further embodiments, R¹⁵ isC₁ haloalkyl. In yet further embodiments, R¹⁵ is CF₃.

In some embodiments of the compound of Formula IIb or pharmaceuticallyacceptable salt thereof, R¹⁴ and R¹⁵ are each independently halo or C₁₋₃haloalkyl (In some further embodiments, each of R¹², R¹³, and R¹⁶ isH.). In some further embodiments, R¹⁴ and R¹⁵ are each independentlyhalo or C₁ haloalkyl. In yet further embodiments, R¹⁴ and R¹⁵ are eachindependently halo.

In some embodiments, the compound of Formula I is a compound of FormulaIII:

In some embodiments of compounds of Formula III or pharmaceuticallyacceptable salt thereof, m is 1.

In some embodiments of compounds of Formula III or pharmaceuticallyacceptable salt thereof, m is 0.

In some embodiments of compounds of Formula III or pharmaceuticallyacceptable salt thereof, at least one of R², R³, R⁴, R⁵, and R⁶ isselected from OH, C₁₋₆ alkoxy, and C₁₋₆ haloalkoxy.

In some embodiments of compounds of Formula III or pharmaceuticallyacceptable salt thereof, at least two of R², R³, R⁴, R⁵, and R⁶ areindependently selected from OH, C₁₋₆ alkoxy, and C₁₋₆ haloalkoxy.

In some embodiments of compounds of Formula III or pharmaceuticallyacceptable salt thereof, at least one of R¹², R¹³, R¹⁴, R¹⁵, and R¹⁶ isother than H.

In some embodiments of the compound of Formula III or pharmaceuticallyacceptable salt thereof, R¹⁴ and R¹⁵ are each independently halo or C₁₋₃haloalkyl. In some further embodiments, R¹⁴ and R¹⁵ are eachindependently halo or C₁ haloalkyl.

At various places in the present specification, substituents ofcompounds of the invention are disclosed in groups or in ranges. It isspecifically Intended that embodiments the invention include each andevery individual subcombination of the members of such groups andranges. For example, the term “C₁₋₆ alkyl” is specifically intended toindividually disclose methyl (C₁ alkyl), ethyl (C₂ alkyl), C₃ alkyl, C₄alkyl, C₅ alkyl, and C₆ alkyl.

For compounds of the invention in which a variable appears more thanonce, each variable can be a different moiety selected from the Markushgroup defining the variable. For example, where a structure is describedhaving two R groups that are simultaneously present on the samecompound, then the two R groups can represent different moietiesselected from the Markush group defined for R.

It is further appreciated that certain features of the invention, whichare, for clarity, described in the context of separate embodiments, canalso be provided in combination in a single embodiment. Conversely,various features of the invention which are, for brevity, described inthe context of a single embodiment, can also be provided separately orin any suitable subcombination.

The term “n-membered” where n is an integer typically describes thenumber of ring-forming atoms in a moiety where the number ofring-forming atoms is n. For example, pyridine is an example of a6-membered heteroaryl ring and thiophene is an example of a 5-memberedheteroaryl group.

As used herein, the term “alkyl” Is meant to refer to a saturatedhydrocarbon group which is straight-chained or branched. Example alkylgroups include, but are not limited to, methyl (Me), ethyl (Et), propyl(e.g., n-propyl and isopropyl), butyl (e.g., n-butyl, isobutyl,t-butyl), pentyl (e.g., n-pentyl, isopentyl, neopentyl), and the like.An alkyl group can contain from 1 to about 20, from 2 to about 20, from1 to about 10, from 1 to about 8, from 1 to about 6, from 1 to about 4,or from 1 to about 3 carbon atoms. The term “alkylene” refers to adivalent alkyl linking group. An example of alkylene is methylene (CH₂).

As used herein, “alkenyl” refers to an alkyl group having one or moredouble carbon-carbon bonds. Example alkenyl groups include, but are notlimited to, ethenyl, propenyl, cyclohexenyl, and the like. The term“alkenylenyl” refers to a divalent linking alkenyl group.

As used herein, “alkynyl” refers to an alkyl group having one or moretriple carbon-carbon bonds. Example alkynyl groups include, but are notlimited to, ethynyl, propynyl, and the like. The term “alkynylenyl”refers to a divalent linking alkynyl group.

As used herein, “haloalkyl” refers to an alkyl group having one or morehalogen substituents. Example haloalkyl groups include, but are notlimited to, CF₃, C₂F₅, CHF₂, CCl₃, CHCl₂, C₂Cl₅, CH₂CF₃, and the like.

As used herein, “aryl” refers to monocyclic or polycyclic (e.g., having2, 3 or 4 fused rings) aromatic hydrocarbons such as, for example,phenyl, naphthyl, anthracenyl, phenanthrenyl, indanyl, indenyl, and thelike. In some embodiments, aryl groups have from 6 to about 20 carbonatoms. In some embodiments, aryl groups have from 6 to about 10 carbonatoms.

As used herein, “cycloalkyl” refers to non-aromatic cyclic hydrocarbonsincluding cyclized alkyl, alkenyl, and alkynyl groups that contain up to20 ring-forming carbon atoms. Cycloalkyl groups can include mono- orpolycyclic (e.g., having 2, 3 or 4 fused rings) ring systems as well asspiro ring systems. A cycloalkyl group can contain from 3 to about 15,from 3 to about 10, from 3 to about 8, from 3 to about 6, from 4 toabout 6, from 3 to about 5, or from 5 to about 6 ring-forming carbonatoms. Ring-forming carbon atoms of a cycloalkyl group can be optionallysubstituted by oxo or sulfido. Example cycloalkyl groups include, butare not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl,cycloheptyl, cyclopentenyl, cyclohexenyl, cyclohexadienyl,cycloheptatrienyl, norbornyl, norpinyl, norcarnyl, adamantyl, and thelike. Also included in the definition of cycloalkyl are moieties thathave one or more aromatic rings fused (i.e., having a bond in commonwith) to the cycloalkyl ring, for example, benzo or thienyl derivativesof pentane, pentene, hexane, and the like (e.g.,2,3-dihydro-1H-indene-1-yl, or 1H-inden-2(3H)-one-1-yl). Preferably,“cycloalkyl” refers to cyclized alkyl groups that contain up to 20ring-forming carbon atoms. Examples of cycloalkyl preferably Includecyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl,adamantyl, and the like.

As used herein, “heteroaryl” groups refer to an aromatic heterocyclehaving up to 20 ring-forming atoms and having at least one heteroatomring member (ring-forming atom) such as sulfur, oxygen, or nitrogen. Insome embodiments, the heteroaryl group has at least one or moreheteroatom ring-forming atoms each independently selected from sulfur,oxygen, and nitrogen. Heteroaryl groups include monocyclic andpolycyclic (e.g., having 2, 3 or 4 fused rings) systems. Examples ofheteroaryl groups include without limitation, pyridyl, pyrimidinyl,pyrazinyl, pyridazinyl, triazinyl, furyl, quinolyl, Isoquinolyl,thienyl, imidazolyl, thiazolyl, indolyl, pyrryl, oxazolyl, benzofuryl,benzothienyl, benzthiazolyl, isoxazolyl, pyrazolyl, triazolyl,tetrazolyl, indazolyl, 1,2,4-thiadiazolyl, isothiazolyl, benzothienyl,purinyl, carbazolyl, benzimidazolyl, indolinyl, and the like. In someembodiments, the heteroaryl group has from 1 to about 20 carbon atoms,and in further embodiments from about 1 to about 5, from about 1 toabout 4, from about 1 to about 3, from about 1 to about 2, carbon atomsas ring-forming atoms. In some embodiments, the heteroaryl groupcontains 3 to about 14, 3 to about 7, or 5 to 6 ring-forming atoms. Insome embodiments, the heteroaryl group has 1 to about 4, 1 to about 3,or 1 to 2 heteroatoms.

As used herein, “heterocycloalkyl” refers to non-aromatic heterocycleshaving up to 20 ring-forming atoms including cyclized alkyl, alkenyl,and alkynyl groups where one or more of the ring-forming carbon atoms isreplaced by a heteroatom such as an O, N, or S atom. Hetercycloalkylgroups can be mono or polycyclic (e.g., both fused and spiro systems).Example “heterocycloalkyl” groups include morpholino, thiomorpholino,piperazinyl, tetrahydrofuranyl, tetrahydrothienyl,2,3-dihydrobenzofuryl, 1,3-benzodioxole, benzo-1,4-dioxane, piperidinyl,pyrrolidinyl, isoxazolidinyl, isothiazolidinyl, pyrazolidinyl,oxazolidinyl, thiazolidinyl, Imidazolidinyl, pyrrolidin-2-one-3-yl, andthe like. Ring-forming carbon atoms and heteroatoms of aheterocycloalkyl group can be optionally substituted by oxo or sulfido.For example, a ring-forming S atom can be substituted by 1 or 2 oxo[i.e., form a S(O) or S(O)₂]. For another example, a ring-forming C atomcan be substituted by oxo (I.e., form carbonyl). Also included in thedefinition of heterocycloalkyl are moieties that have one or morearomatic rings fused (i.e., having a bond in common with) to thenonaromatic heterocyclic ring, for example pyridinyl, thiophenyl,phthalimidyl, naphthalimidyl, and benzo derivatives of heterocycles suchas indolene, isoindolene, isoindolin-1-one-3-yl,4,5,6,7-tetrahydrothieno[2,3-c]pyridine-5-yl,5,6-dihydrothieno[2,3-c]pyridin-7(4H)-one-5-yl, and3,4-dihydroisoquinolin-1 (2H)-one-3yl groups. Ring-forming carbon atomsand heteroatoms of the heterocycloalkyl group can be optionallysubstituted by oxo or sulfido. In some embodiments, the heterocycloalkylgroup has from 1 to about 20 carbon atoms, and in further embodimentsfrom about 3 to about 20 carbon atoms. In some embodiments, theheterocycloalkyl group contains 3 to about 14, 3 to about 7, or 5 to 6ring-forming atoms. In some embodiments, the heterocycloalkyl group has1 to about 4, 1 to about 3, or 1 to 2 heteroatoms. In some embodiments,the heterocycloalkyl group contains 0 to 3 double bonds. In someembodiments, the heterocycloalkyl group contains 0 to 2 triple bonds.

As used herein, “halo” or “halogen” includes fluoro, chloro, bromo, andiodo.

As used herein, “alkoxy” refers to an —O-alkyl group. Example alkoxygroups include methoxy, ethoxy, propoxy (e.g., n-propoxy andisopropoxy), t-butoxy, and the like.

As used herein, “haloalkoxy” refers to an —O-haloalkyl group. An examplehaloalkoxy group is OCF₃. As used herein, “trihalomethoxy” refers to amethoxy group having three halogen substituents. Examples oftrihalomethoxy groups include, but are not limited to, —OCF₃, —OCClF₂,—OCCl₃, and the like.

As used herein, “arylalkyl” refers to a C₁₋₆ alkyl substituted by aryland “cycloalkylalkyl” refers to C₁₋₆ alkyl substituted by cycloalkyl.

As used herein, “heteroarylalkyl” refers to a C₁₋₈ alkyl groupsubstituted by a heteroaryl group, and “heterocycloalkylalkyl” refers toa C₁₋₆ alkyl substituted by heterocycloalkyl.

As used herein, “amino” refers to NH₂.

As used herein, “alkylamino” refers to an amino group substituted by analkyl group.

As used herein, “dialkylamino” refers to an amino group substituted bytwo alkyl groups.

As used here, C(O) refers to C(═O).

As used here, C(S) refers to C(═S).

As used here, S(O) refers to S(═O).

As used here, S(O)₂ refers to S(═O)₂.

As used herein, the term “optionally substituted” means thatsubstitution is optional and therefore includes both unsubstituted andsubstituted atoms and moieties. A “substituted” atom or moiety indicatesthat any hydrogen on the designated atom or moiety can be replaced witha selection from the indicated substituent group, provided that thenormal valency of the designated atom or moiety is not exceeded, andthat the substitution results in a stable compound. For example, if amethyl group (i.e., CH₃) is optionally substituted, then 3 hydrogenatoms on the carbon atom can be replaced with substituent groups.

As used herein, “about” in connection with a numerical value means thatthe numerical value is approximate and small variations would notsignificantly affect the practice of the disclosed embodiments. Where anumerical limitation is used, unless indicated otherwise by the context,“about” means the numerical value can vary by ±10% and remain within thescope of the disclosed embodiments.

The compounds described in the embodiments herein can be asymmetric(e.g., having one or more stereocenters). All stereoisomers, such asenantiomers and diastereomers, are intended unless otherwise indicated.Compounds of the present invention that contain asymmetricallysubstituted carbon atoms can be isolated in optically active or racemicforms. Methods on how to prepare optically active forms from opticallyactive starting materials are known in the art, such as by resolution ofracemic mixtures or by stereoselective synthesis. Many geometric isomersof olefins, C═N double bonds, and the like can also be present in thecompounds described herein, and all such stable isomers are contemplatedin the present invention. Cis and trans geometric isomers of thecompounds of the present invention are described and may be Isolated asa mixture of isomers or as separated isomeric forms. Where a compoundcapable of stereoisomerism or geometric isomerism is designated in itsstructure or name without reference to specific R/S or cis/transconfigurations, it is intended that all such isomers are contemplated.

Resolution of racemic mixtures of compounds can be carried out by any ofnumerous methods known in the art. An example method includes fractionalrecrystallization using a chiral resolving acid which is an opticallyactive, salt-forming organic acid. Suitable resolving agents forfractional recrystallization methods are, for example, optically activeacids, such as the D and L forms of tartaric acid, diacetyltartaricacid, dibenzoyltartaric acid, mandelic acid, malic acid, lactic acid orthe various optically active camphorsulfonic acids such as3-camphorsulfonic acid. Other resolving agents suitable for fractionalcrystallization methods include stereoisomerically pure forms ofα-methylbenzylamine (e.g., Sand R forms, or diastereomerically pureforms), 2-phenylglycinol, norephedrine, ephedrine, N-methylephedrine,cyclohexylethylamine, 1,2-diaminocyclohexane, and the like.

Resolution of racemic mixtures can also be carried out by elution on acolumn packed with an optically active resolving agent (e.g.,dinitrobenzoylphenylglycine). Suitable elution solvent composition canbe determined by one skilled in the art.

Compounds of embodiments the invention also include tautomeric forms.Tautomeric forms result from the swapping of a single bond with anadjacent double bond together with the concomitant migration of aproton. Tautomeric forms include prototropic tautomers which areisomeric protonation states having the same empirical formula and totalcharge. Example prototropic tautomers include ketone—enol pairs,amide—imidic acid pairs, lactam—lactim pairs, amide—imdic acid pairs,enamine—Imine pairs, and annular forms where a proton can occupy two ormore positions of a heterocyclic system, for example, 1H- and3H-imidazole, 1H-, 2H- and 4H-1,2,4-triazole, 1H- and 2H-isoindole, and1H- and 2H-pyrazole. Tautomeric forms can be in equilibrium orsterically locked into one form by appropriate substitution.

Compounds of embodiments the invention further include hydrates andsolvates, as well as anhydrous and non-solvated forms.

The term, “compound,” as used herein is meant to include allstereoisomers, geometric iosomers, tautomers, and isotopes of thestructures depicted.

All compounds and pharmaceutically acceptable salts thereof, can beprepared or present together with other substances such as water andsolvents (e.g. hydrates and solvates) or can be isolated.

Compounds of embodiments the invention can also include all isotopes ofatoms occurring in the intermediates or final compounds. IsotopesInclude those atoms having the same atomic number but different massnumbers. For example, isotopes of hydrogen include tritium anddeuterium.

In some embodiments, the compounds of the invention, or salts thereof,are substantially isolated. By “substantially isolated” is meant thatthe compound is at least partially or substantially separated from theenvironment in which is formed or detected. Partial separation caninclude, for example, a composition enriched in the compound of theinvention. Substantial separation can include compositions containing atleast about 50%, at least about 60%, at least about 70%, at least about80%, at least about 90%, at least about 95%, at least about 97%, or atleast about 99% by weight of the compound of the invention, or saltthereof. Methods for isolating compounds and their salts are routine inthe art.

Compounds of embodiments the invention are Intended to Include compoundswith stable structures. As used herein, “stable compound” and “stablestructure” are meant to indicate a compound that is sufficiently robustto survive isolation to a useful degree of purity from a reactionmixture, and formulation into an efficacious therapeutic agent.

The phrase “pharmaceutically acceptable” is employed herein to refer tothose compounds, materials, compositions, and/or dosage forms which are,within the scope of sound medical judgment, suitable for use in contactwith the tissues of human beings and animals without excessive toxicity,irritation, allergic response, or other problem or complication,commensurate with a reasonable benefit/risk ratio.

The expressions, “ambient temperature” and “room temperature,” as usedherein, are understood in the art, and refer generally to a temperature,e.g. a reaction temperature, that is about the temperature of the roomin which the reaction Is carried out, for example, a temperature fromabout 20° C. to about 30° C.

Embodiments of the present invention also includes pharmaceuticallyacceptable salts of the compounds described herein. As used herein,“pharmaceutically acceptable salts” refers to derivatives of thedisclosed compounds wherein the parent compound is modified byconverting an existing acid or base moiety to its salt form. Examples ofpharmaceutically acceptable salts include, but are not limited to,mineral or organic acid salts of basic residues such as amines; alkalior organic salts of acidic residues such as carboxylic acids; and thelike. The pharmaceutically acceptable salts of the present Inventioninclude the conventional non-toxic salts of the parent compound formed,for example, from non-toxic inorganic or organic acids. Thepharmaceutically acceptable salts of the present invention can besynthesized from the parent compound which contains a basic or acidicmoiety by conventional chemical methods. Generally, such salts can beprepared by reacting the free acid or base forms of these compounds witha stoichiometric amount of the appropriate base or acid in water or inan organic solvent, or in a mixture of the two; generally, nonaqueousmedia like ether, ethyl acetate, ethanol, isopropanol, or acetonitrile(ACN) are preferred. Lists of suitable salts are found in Remington'sPharmaceutical Sciences, 17th ed., Mack Publishing Company, Easton, Pa.,1985, p. 1418 and Journal of Pharmaceutical Science, 66, 2 (1977), eachof which is incorporated herein by reference in its entirety.

Synthesis

Compounds of embodiments the invention, including salts thereof, can beprepared using known organic synthesis techniques and can be synthesizedaccording to any of numerous possible synthetic routes.

The reactions for preparing compounds of the Invention can be carriedout in suitable solvents which can be readily selected by one of skillin the art of organic synthesis. Suitable solvents can be substantiallynon-reactive with the starting materials (reactants), the intermediates,or products at the temperatures at which the reactions are carried out,e.g., temperatures which can range from the solvent's freezingtemperature to the solvent's boiling temperature. A given reaction canbe carried out in one solvent or a mixture of more than one solvent.Depending on the particular reaction step, suitable solvents for aparticular reaction step can be selected by the skilled artisan.

Preparation of compounds of the invention can involve the protection anddeprotection of various chemical groups. The need for protection anddeprotection, and the selection of appropriate protecting groups, can bereadily determined by one skilled in the art. The chemistry ofprotecting groups can be found, for example, in T. W. Greene and P. G.M. Wuts, Protective Groups in Organic Synthesis, 3^(rd) Ed., Wiley &Sons, Inc., New York (1999), which is incorporated herein by referencein its entirety.

Reactions can be monitored according to any suitable method known in theart. For example, product formation can be monitored by spectroscopicmeans, such as nuclear magnetic resonance spectroscopy (e.g., ¹H or¹³C), infrared spectroscopy, spectrophotometry (e.g., UV-visible), massspectrometry, or by chromatographic methods such as high performanceliquid chromatography (HPLC) or thin layer chromatography (TLC).

The compounds of embodiments of the invention can be prepared, forexample, according to the reaction pathways, synthetic procedures, andtechniques described below.

As shown in Scheme 1, benzaldehyde derivative 1-1 can be reacted withacetaldehyde in the presence of either an acid or a base catalyst toafford cinnamic aldehyde 1-2. Reaction of cinnamic aldehyde 1-2 with anorganometallic compound such as a Grignard reagent R⁸MgX¹ [wherein X¹ ishalo such as Cl or Br], followed by oxidation of the intermediatealcohol to ketone and by reduction of the C═C bond to a C—C single bondunder a hydrogenation condition (for example, in the presence of Pd/Ccatalyst), affords ketone 1-3. Reaction of ketone 1-3 with amine 1-4under reductive amination condition (such as in the presence of aborohydride reducing reagent) affords compound 1-5.

As shown in Scheme 2, Ketone 2-1 can be reacted with an organometalliccompound such as a Grignard reagent R⁷MgX² [wherein R⁷ can be C₁₋₆ alkylor C₃₋₇cycloalkyl; and X² can be halo such as Cl or Br] to affordalcohol 2-2a. Reduction of Ketone 2-1 such as in the presence of aborohydride reducing reagent affords alcohol 2-2b. The OH group ofalcohol 2-2 (wherein R⁷ can be H, C₁₋₆ alkyl, or C₃₋₇ cycloalkyl) can beconverted to a better leaving group Lg¹ such as OMs [mesylate orCH₃S(O)₂O—] or OTf [triflate or CF₃S(O)₂O—], followed by reaction withNH₃ to afford amine 2-4. Ketone 2-1 can also undergo reductive aminationwith NH₃ to afford amine 2-5.

As shown in Scheme 3, amine 3-1 (wherein R⁷ can be, e.g., H, C₁₋₆ alkyl,or C₃₋₇ cycloalkyl) can be reacted with compound 3-2 [Lg² can be aleaving group such as triflate group (—OTf) or halo (e.g. Cl or Br)] toafford compound 3-3.

As shown in Scheme 4, benzaldehyde derivative 4-1 can be reacted with amethylmetalic compound such as a Grignard reagent MeMgX¹ [wherein X¹ ishalo such as CI or Br], followed by oxidation of the intermediatealcohol to ketone to afford ketone 4-2. Ester compound 4-3 [wherein R′can be alkyl (e.g. methyl or ethyl) or arylalkyl; and Pg¹ can be anamine protecting group (such as tert-butyloxycarbonyl or Boc;benzyloxycarbonyl or Cbz; or benzyl)] can be reduced to an alcohol (inthe presence of a reducing reagent such as Lithium aluminium hydride orLAH), followed by conversion of the OH group to a better leaving groupLg³ such as OMs [mesylate or CH₃S(O)₂O—] or OTf [triflate orCF₃S(O)₂O—], to afford compound 4-4.

Reaction of compound 4-2 with compound 4-4 in the presence of a strongbase (such as lithium diisopropylamide or LDA), followed byhydrogenation (such as in the presence of a Pd/C catalyst) to reduce theC(O) to CH₂ and by removal of the protecting group Pg¹ under suitableconditions (for example, a benzyl group can be removed underhydrogenation condition in the presence of Pd/C; or Boc group can beremoved under acidic condition), affords amine 4-5. Amine 4-5 can bereacted with compound 4-6 [wherein Lg² can be a leaving group such astriflate group (—OTf) or halo (e.g. Cl or Br)] to afford compound 4-7.

Those skilled in the art can recognize that in all of the schemesdescribed herein, if there are functional (reactive) groups present on asubstituent group such as R¹, R², R³, R⁴, R⁵, R⁶, R⁷, R⁸, R⁹, R¹⁰, R¹¹,R¹², R¹³, R¹⁴, R¹⁵, R¹⁶, etc., further modification can be made ifappropriate and/or desired. For example, a CN group can be hydrolyzed toafford an amide group; a carboxylic acid can be converted to an amide; acarboxylic acid can be converted to an ester, which in turn can bereduced to an alcohol, which in turn can be further modified. Foranother example, an OH group can be converted into a better leavinggroup such as mesylate, which in turn is suitable for nucleophilicsubstitution, such as by CN. For another example, an —S— can be oxidizedto —S(O)— and/or —S(O)₂—. For yet another example, unsaturated bond suchas C═C or C≡C can be reduced to saturated bond by hydrogenation. In someembodiments, a primary amine or a secondary amine moiety (present on asubstituent group such as R¹-R¹⁶, etc.) can be converted to amide,sulfonamide, urea, or thiourea moiety by reacting it with an appropriatereagent such as an acid chloride, a sulfonyl chloride, an isocyanate, ora thioisocyanate compound. Thus, a compound of Formula I (such ascompound 3-3 of Scheme 3) having a substituent which contains afunctional group can be converted to another compound of Formula Ihaving a different substituent group.

As used herein, the term “reacting” refers to the bringing together ofdesignated chemical reactants such that a chemical transformation takesplace generating a compound different from any initially introduced intothe system. Reacting can take place in the presence or absence ofsolvent.

Chemical Conditioning

In some embodiments, a method of preparing an array of chemicalcompounds from a biological extract such as ginger oil is provided.

The method of the invention, termed “chemical conditioning” is generallyapplicable to all biological extracts, in particular, natural plantextracts, common or medicinal. See e.g. US20080193574 and WO2008042755,each of which is incorporated herein by reference in its entirety.Chemical conditioning is a method which produces novel unnaturaldrug-like compounds from readily available natural materials. Ingeneral, the “chemical conditioning” of natural extracts coupled withpre-fractionation of the chemically conditioned extracts facilitatessuccessful biochemical screening of extracts by destroying reactivenatural compounds that generate false positive results in biochemicalassays. Chemical conditioning produces novel lead-like and drug-likecompounds and, the reductive amination protocol described here canproduce structurally diverse nitrogen-containing products that areparticularly lead-like and drug-like.

In certain embodiments of the present invention, a method of preparingchemical compounds from a biological extract is exemplified in Scheme 5abelow. According to the method, first, a biological extract, e.g., aplant extract is provided, the biological extract has one or morebiological compounds, each biological compound having one or morereactive electrophilic groups. Next, the biological compounds in thebiological extract are reacted with an amine to incorporate the amineinto the biological compounds. Next, the biological compounds having theincorporated amine are reacted with a reducing agent to reduce theintermediate imine and enamine compounds and form one or morenitrogen-containing chemical compounds. Thus, the resultantnitrogen-containing chemical compounds are derivatives of the biologicalcompounds in the biological extract. In some embodiments, the biologicalcompounds in the biological extract are compounds having ketones andaldehydes that are reacted with various amines. This reaction isfollowed by hydride reduction of the intermediate imines and enamines toprovide secondary and tertiary amines. The reaction of ketones andaldehydes with amines, followed by reduction to form imines and enaminesis known in the art.

R′ and R″ represent a variety of substituents that make up a biologicalcompound; andR* represents a variety of substituent(s) that, together with thenitrogen, make up an amine compound.

The chemical conditioning method described herein employs a biologicalextract, using many different reagents, to efficiently produce an arrayof nitrogen-containing chemical compounds. The ready commercialavailability of many low molecular weight amines for use as inputs inthe reductive amination sequence enables the development of manydifferent and structurally diverse central nervous system druglikemixtures from the same natural extract. Suitable amines for use in thepresent method are selected from the group consisting of primary amines,secondary amines, cyclic amines, pyrollidine, and amino acids. Suitablereducing agents for use in the present method are selected from thegroup of hydride reducing agents including but not limited to sodiumborohydride, sodium triacetoxyborohydride, and lithium aluminum hydride.

The method may further comprise quenching the reaction a quenchingagent, wherein the quenching agent is selected from but not limited tothe group consisting of sodium bicarbonate, sodium carbonate, sodiumsulfate, sodium sulfate decahydrate. The method may also furthercomprise isolating one or more of the nitrogen-containing chemicalcompounds, in a purified or unpurified form. The resultantnitrogen-containing chemical may then be screened or tested forbiological activity.

The process of chemical conditioning by reductive amination describedherein destroys reactive electrophiles in the natural extract, includingketones, as in the gingerols, and converts them to chemically stablecompounds such as amines. The resulting conditioned extracts containboth natural compounds and novel unnatural nitrogen-containing amineproducts that are potential drug candidates. In the case of the extractsof gingerol, the nitrogen-containing amine products are potentialcentral nervous system drugs.

For the purpose of this disclosure, the following terms have thefollowing meanings.

The term “biological compound” as used herein refers to a chemicalcompound that occurs in nature.

The term “biological extract” as used herein refers to an extract from abiological sample, such as a plant extract, or other extract fromorganic matter, containing chemical compounds that occur in nature.

The term “reactive electrophilic group” as used herein refers to an atomor group of atoms that has the ability to react with a nucleophile.

The term “nitrogen-containing derivative” as used herein representsthose derivatives containing a nitrogen atom, where the nitrogen atom isa substitution another atom, such as oxygen in the parent compound.

In one embodiment, a specific example of the chemical conditioningprocess is shown in Scheme 5 below. Scheme 5 shows the two-stepreductive amination chemical conditioning protocol performed on gingeroil and ginger oleoresin in accordance with one embodiment of themethod, wherein ginger oil or gingerol comprising ketone 5-1 areconverted to amine 5-4. According to the method shown in Scheme 5,ginger oil (an extract of ginger containing ketone 5-1 and othermolecules occurring in natural ginger) is reacted with amine 5-2 to formcompound 5-3. Then, the resultant compound 5-3 is then reduced, with areducing agent such as a borohydride, to from the nitrogen-containingcompound 5-4 (the reaction crude product also include other chemicalcompounds).

In the next step of the method, amine 5-4 is isolated/purified from theextract (the crude reaction product of the 2-step reductive amination).The conditioned extracts can be fractionated by flash chromatography.The fraction that contains amine 5-4 can undergo furtherpurification/isolation according to the methods known to those in theart. Further isolation and characterization of the fraction thatcontains amine 5-4 may follow. The isolated amine 5-4 is tested for itsbiological activity such as by those methods described hereinwith.

Some examples of benzylamine 5-2 used in the chemical conditioningprocess of the invention shown in Scheme 5 include:

New lead compounds generated by this chemical conditioning method canalso be prepared to the synthetic methods described hereinwith.

In some embodiments, the derivatives of ginger oil such as amine 5-4possess beta-secretase inhibitory activity, and/or inhibit amyloidproduction, amyloid assembly, the activity/effect of Abeta oligomers onneurons (such as neurons in the brain), amyloid aggregation, amyloid(including amyloid oligomer) binding, or amyloid deposition. Thesecompounds are useful therapeutic agents for the treatment and preventionof cognitive decline, amyloid production, neurodegeneration, andAlzhelmer's disease.

Methods

In some embodiments, the compounds of present invention inhibit, treat,or abate (partially inhibit) binding of amyloid (including Abetaoligomers) to neurons (such as neurons in the brain) and are useful forthe inhibition, treatment, and abatement of cognitive decline and/orAlzheimer's disease. In some embodiments, the compounds of presentinvention inhibit, treat, or abate (partially inhibit) one or more ofamyloid aggregation, amyloid oligomer binding, and amyloid deposition.In some embodiments, the compounds of present invention inhibit, treat,or abate (partially inhibit) amyloid deposition. In some embodiments,the compounds of present invention inhibit, treat, or abate (partiallyinhibit) the activity/effect of Abeta oligomers on neurons (such asneurons in the brain) and are useful for the inhibition, treatment, andabatement of cognitive decline and/or Alzheimer's disease. In someembodiments, the compounds of present invention inhibit, treat, or abate(partially inhibit) the activity/effect of Abeta oligomers on neurons(such as neurons in the brain) via disruption of Abeta oligomers,inhibition of Abeta oligomer binding to neurons, and/or counteraction ofsignal transduction mechanisms of action initiated by Abeta oligomerbinding.

In some embodiments, the compounds show activity in a beta-secretaseassay and are useful for the inhibition, treatment, and abatement ofcognitive decline and Alzheimer's disease. In some embodiments thederivative of ginger oil is a compound in purified and Isolated form(for example, with a purity of greater than 80%, 85%, 90%, 95%, 98%, or99% by weight). The compounds and methods described herein may be usedto treat one or more symptoms of cognitive decline and/or Alzheimer'sdisease such as memory loss, confusion, Impaired judgment, personalitychanges, disorientation, and loss of language skills. Further, thecompounds and methods described herein may be useful in inhibiting,treating, and/or abating cognitive decline and/or Alzheimer's disease byrestoring long term potentiation, and/or inhibiting, treating, orabatement of one or both of neurodegeneration and general amyloidosis,more specifically, by inhibiting, treating, or abatement of one or moreof amyloid production, amyloid assembly, amyloid aggregation, amyloid(including amyloid oligomer) binding, and amyloid deposition.

In some embodiments, compounds of the invention can inhibit, treat, orabate one or more of amyloid production, amyloid assembly, amyloidaggregation, amyloid oligomer binding, and amyloid deposition. In someembodiments, compounds of the invention can restore long termpotentiation, inhibit, treat, or abate one or both of neurodegenerationand general amyloidosis.

In some embodiments, compounds of present invention inhibit, treat, orabate (partially inhibit) one or more of amyloid aggregation, amyloidoligomer binding, and amyloid deposition. In some embodiments, thecompounds of present invention inhibit (or partially inhibit) amyloiddeposition. In some embodiments, the compounds of present inventionInhibit, treat, or abate (partially inhibit) binding of amyloid(including Abeta oligomers) to neurons (such as neurons in the brain).In some embodiments, the compounds of present invention are useful forthe inhibition, treatment, and abatement of cognitive decline and/orAlzheimer's disease.

In some embodiments, compounds of the invention can inhibit activity ofbeta-secretase. In some embodiments, compounds of the invention can beused in methods of inhibiting activity of beta-secretase by contactingthe beta-secretase with any one or more of the compounds or compositionsdescribed herein.

Another aspect of the present invention pertains to methods of treatingcognitive decline and/or Alzheimer's disease in an individual (e.g.,patient) by administering to the individual a therapeutically effectiveamount or dose of a compound of the present invention or apharmaceutical composition thereof.

Treatment of the diseases/disorders herein includes treating one or moresymptoms associated with the diseases/disorders, for example, symptomsof cognitive decline and/or Alzheimer's disease.

As used herein, the term “contacting” refers to the bringing together ofindicated moieties in an in vitro system or an in vivo system. Forexample, “contacting” a beta-secretase or a neuron cell (or a neuroncell in the presence of one or more of beta-amyloid oligomers) with acompound of the invention includes the administration of a compound ofthe present invention to an individual or patient, such as a human,having a beta-secretase or a neuron cell, as well as, for example,introducing a compound of the invention into a sample containing acellular or purified preparation containing the a beta-secretase or aneuron cell (or a neuron cell in the presence of one or more ofbeta-amyloid oligomers).

As used herein, the term “individual” or “patient,” usedinterchangeably, refers to any animal, including mammals, preferablymice, rats, other rodents, rabbits, dogs, cats, swine, cattle, sheep,horses, or primates, and most preferably humans.

As used herein, the phrase “therapeutically effective amount” refers tothe amount of active compound or pharmaceutical agent that elicits thebiological or medicinal response that is being sought in a tissue,system, animal, individual or human by a researcher, veterinarian,medical doctor or other clinician.

As used herein, the term “treating” or “treatment” refers to one or moreof (1) preventing the disease; for example, preventing a disease,condition or disorder in an individual who may be predisposed to thedisease, condition or disorder but does not yet experience or displaythe pathology or symptomotology of the disease; (2) inhibiting/retardingthe disease; for example, inhibiting/retarding a disease, condition ordisorder in an individual who is experiencing or displaying thepathology or symptomotology of the disease, condition or disorder; and(3) ameliorating the disease; for example, ameliorating a disease,condition or disorder in an individual who is experiencing or displayingthe pathology or symptomotology of the disease, condition or disorder(i.e., reversing the pathology and/or symptomotology) such as decreasingthe severity of disease or completely eliminating/curing the disease. Asused herein, treating a disease further includes treating one or moresymptoms associated with the disease.

Combination Therapies

In certain embodiments, one or more additional pharmaceutical agents fortreatment of cognitive decline and/or Alzheimer's disease can be used incombination with the compounds of the present invention for treatment ofcognitive decline and/or Alzheimer's disease. The one or more additionalpharmaceutical agents can be administered to a patient simultaneously orsequentially.

Pharmaceutical Formulations and Dosage Forms

In certain embodiments, the compounds of the Invention can beadministered in the form of pharmaceutical compositions. Thesecompositions can be prepared in a manner well known in thepharmaceutical art, and can be administered by a variety of routes,depending upon whether local or systemic treatment Is desired and uponthe area to be treated. Administration may be topical (includingtransdermal, epidermal, ophthalmic and to mucous membranes includingintranasal, vaginal and rectal delivery), pulmonary (e.g., by inhalationor insufflation of powders or aerosols, including by nebulizer;intratracheal or intranasal), oral or parenteral. Parenteraladministration includes intravenous, intraarterial, subcutaneous,intraperitoneal intramuscular or injection or infusion; or intracranial,e.g., intrathecal or intraventricular, administration. Parenteraladministration can be in the form of a single bolus dose, or may be, forexample, by a continuous perfusion pump. Pharmaceutical compositions andformulations for topical administration may include transdermal patches,ointments, lotions, creams, gels, drops, suppositories, sprays, liquidsand powders. Conventional pharmaceutical carriers, aqueous, powder oroily bases, thickeners and the like may be necessary or desirable.Coated condoms, gloves and the like may also be useful.

Embodiments of this invention also include pharmaceutical compositionswhich contain, as the active ingredient, one or more of the compounds ofthe invention above in combination with one or more pharmaceuticallyacceptable carriers (excipients). In making the compositions of theInvention, the active ingredient is typically mixed with an excipient,diluted by an excipient or enclosed within such a carrier in the formof, for example, a capsule, sachet, paper, or other container. When theexcipient serves as a diluent, it can be a solid, semi-solid, or liquidmaterial, which acts as a vehicle, carrier or medium for the activeingredient. Thus, the compositions can be in the form of tablets, pills,powders, lozenges, sachets, cachets, elixirs, suspensions, emulsions,solutions, syrups, aerosols (as a solid or in a liquid medium),ointments containing, for example, up to 10% by weight of the activecompound, soft and hard gelatin capsules, suppositories, sterileinjectable solutions, and sterile packaged powders.

In preparing a formulation, the active compound can be milled to providethe appropriate particle size prior to combining with the otheringredients. If the active compound is substantially insoluble, it canbe milled to a particle size of less than 200 mesh. If the activecompound is substantially water soluble, the particle size can beadjusted by milling to provide a substantially uniform distribution inthe formulation, e.g. about 40 mesh.

The compounds of the invention may be milled using known millingprocedures such as wet milling to obtain a particle size appropriate fortablet formation and for other formulation types. Finely divided (nanoparticulate) preparations of the compounds of the invention can beprepared by processes known in the art, for example see InternationalPatent Application No. WO 2002/000196.

Some examples of suitable excipients include lactose, dextrose, sucrose,sorbitol, mannitol, starches, gum acacia, calcium phosphate, alginates,tragacanth, gelatin, calcium silicate, microcrystalline cellulose,polyvinylpyrrolidone, cellulose, water, syrup, and methyl cellulose. Theformulations can additionally include: lubricating agents such as talc,magnesium stearate, and mineral oil; wetting agents; emulsifying andsuspending agents; preserving agents such as methyl- andpropylhydroxy-benzoates; sweetening agents; and flavoring agents. Thecompositions of the invention can be formulated so as to provide quick,sustained or delayed release of the active ingredient afteradministration to the patient by employing procedures known in the art.

The compositions can be formulated in a unit dosage form, each dosagecontaining from about 5 to about 1000 mg (1 g), more usually about 100to about 500 mg, of the active ingredient. The term “unit dosage forms”refers to physically discrete units suitable as unitary dosages forhuman subjects and other mammals, each unit containing a predeterminedquantity of active material calculated to produce the desiredtherapeutic effect, in association with a suitable pharmaceuticalexcipient.

The active compound can be effective over a wide dosage range and can begenerally administered in a pharmaceutically effective amount. Forexample, the dosage of the active compounds of the invention as employedfor the treatment of a patient in need thereof (such as an adult human)may range from 0.1 to 3000 mg per day, depending on the route andfrequency of administration. Such a dosage corresponds to 0.001 to 50mg/kg per day. In some embodiments, the dosage of the active compoundsof the invention as employed for the treatment of a patient in needthereof (such as an adult human) may range from 1 to 2000 mg per day,from 1 to 1000 mg per day, from 10 to 1000 mg per day, or from to 500 mgper day. It will be understood, however, that the amount of the compoundactually administered will usually be determined by a physician,according to the relevant circumstances, including the condition to betreated, the chosen route of administration, the actual compoundadministered, the age, weight, and response of the individual patient,the severity of the patient's symptoms, and the like.

For preparing solid compositions such as tablets, the principal activeingredient can be mixed with a pharmaceutical excipient to form a solidpre-formulation composition containing a homogeneous mixture of acompound of the present invention. When referring to thesepre-formulation compositions as homogeneous, the active ingredient istypically dispersed evenly throughout the composition so that thecomposition can be readily subdivided into equally effective unit dosageforms such as tablets, pills and capsules. This solid pre-formulation isthen subdivided into unit dosage forms of the type described abovecontaining from, for example, about 0.1 to about 1000 mg of the activeingredient of the present invention.

The tablets or pills of the present invention can be coated or otherwisecompounded to provide a dosage form affording the advantage of prolongedaction. For example, the tablet or pill can comprise an inner dosage andan outer dosage component, the latter being in the form of an envelopeover the former. The two components can be separated by an enteric layerwhich serves to resist disintegration in the stomach and permit theinner component to pass intact into the duodenum or to be delayed inrelease. A variety of materials can be used for such enteric layers orcoatings, such materials including a number of polymeric acids andmixtures of polymeric acids with such materials as shellac, cetylalcohol, and cellulose acetate.

The liquid forms in which the compounds and compositions of the presentinvention can be incorporated for administration orally or by injectioninclude aqueous solutions, suitably flavored syrups, aqueous or oilsuspensions, and flavored emulsions with edible oils such as cottonseedoil, sesame oil, coconut oil, or peanut oil, as well as elixirs andsimilar pharmaceutical vehicles.

Compositions for inhalation or insufflation include solutions andsuspensions in pharmaceutically acceptable, aqueous or organic solvents,or mixtures thereof, and powders. The liquid or solid compositions maycontain suitable pharmaceutically acceptable excipients as describedsupra. In some embodiments, the compositions are administered by theoral or nasal respiratory route for local or systemic effect.Compositions in can be nebulized by use of inert gases. Nebulizedsolutions may be breathed directly from the nebulizing device or thenebulizing device can be attached to a face masks tent, or intermittentpositive pressure breathing machine. Solution, suspension, or powdercompositions can be administered orally or nasally from devices whichdeliver the formulation in an appropriate manner.

The amount of compound or composition administered to a patient willvary depending upon what is being administered, the purpose of theadministration, such as prophylaxis or therapy, the state of thepatient, the manner of administration, and the like. In therapeuticapplications, compositions can be administered to a patient alreadysuffering from a disease in an amount sufficient to cure or at leastpartially arrest the symptoms of the disease and its complications.Effective doses will depend on the disease condition being treated aswell as by the judgment of the attending clinician depending uponfactors such as the severity of the disease, the age, weight and generalcondition of the patient, and the like.

The compositions administered to a patient can be in the form ofpharmaceutical compositions described above. These compositions can besterilized by conventional sterilization techniques, or may be sterilefiltered. Aqueous solutions can be packaged for use as is, orlyophilized, the lyophilized preparation being combined with a sterileaqueous carrier prior to administration. The pH of the compoundpreparations typically will be between 3 and 11, more preferably from 5to 9 and most preferably from 7 to 8. It will be understood that use ofcertain of the foregoing excipients, carriers, or stabilizers willresult in the formation of pharmaceutical salts.

The therapeutic dosage of the compounds of the present invention canvary according to, for example, the particular use for which thetreatment is made, the manner of administration of the compound, thehealth and condition of the patient, and the judgment of the prescribingphysician. The proportion or concentration of a compound of theinvention in a pharmaceutical composition can vary depending upon anumber of factors including dosage, chemical characteristics (e.g.,hydrophobicity), and the route of administration. For example, thecompounds of the invention can be provided in an aqueous physiologicalbuffer solution containing about 0.1 to about 10% w/v of the compoundfor parenteral administration. Some typical dose ranges are from about 1g/kg to about 1 g/kg of body weight per day. In some embodiments, thedose range is from about 0.01 mg/kg to about 100 mg/kg of body weightper day. The dosage is likely to depend on such variables as the typeand extent of progression of the disease or disorder, the overall healthstatus of the particular patient, the relative biological efficacy ofthe compound selected, formulation of the excipient, and its route ofadministration. Effective doses can be extrapolated from dose-responsecurves derived from in vitro or animal model test systems.

The compositions of the invention can further include one or moreadditional pharmaceutical agents such as a chemotherapeutic, steroid,anti-inflammatory compound, or immunosuppressant, examples of which arelisted hereinabove.

Labeled Compounds and Assay Methods

Another aspect of the present invention relates to labeled compounds ofthe invention (radio-labeled, fluorescent-labeled, etc.) that would beuseful not only in radio-imaging but also in assays, both in vitro andin vivo, for localizing and quantitating the enzyme in tissue samples,including human, and for identifying ligands by inhibition binding of alabeled compound. Accordingly, the present invention includes enzymeassays that contain such labeled compounds.

Embodiments of the present invention further includesisotopically-labeled compounds of the invention. An “isotopically” or“radio-labeled” compound is a compound of the invention where one ormore atoms are replaced or substituted by an atom having an atomic massor mass number different from the atomic mass or mass number typicallyfound in nature (i.e., naturally occurring). Suitable radionuclides thatmay be incorporated in compounds of the present invention include butare not limited to ²H (also written as D for deuterium), ³H (alsowritten as T for tritium), ¹¹C, ¹³C, ¹⁴C, ¹³N, ¹⁵N ¹⁵O, ¹⁷O, ¹⁸O, ¹⁸F,³⁵S, ³⁶Cl, ⁸²Br, ⁷⁵Br, ⁷⁶Br, ⁷⁷Br, ¹²³I, ¹²⁴I, ¹²⁵I and ¹³¹I. Theradionuclide that is incorporated in the radio-labeled compounds willdepend on the specific application of that radio-labeled compound. Forexample, for in vitro receptor labeling and competition assays,compounds that incorporate ³H, ¹⁴C, ⁸²Br, ¹²⁵I, ¹³¹I, ³⁵S or willgenerally be most useful. For radio-imaging applications ¹¹C, ¹⁸F, ¹²⁵I,¹²³I, ¹²⁴I, ¹³¹I, ⁷⁵Br, ⁷⁶Br or ⁷⁷Br will generally be most useful.

It is understood that a “radio-labeled compound” is a compound that hasincorporated at least one radionuclide. In some embodiments theradionuclide is selected from ³H, ¹⁴C, ¹²⁵I, ³⁵S and ⁸²Br.

In some embodiments, the labeled compounds of the present inventioncontain a fluorescent label.

Synthetic methods for incorporating radio-isotopes and fluorescentlabels into organic compounds are well known in the art.

A labeled compound of the invention (radio-labeled, fluorescent-labeled,etc.) can be used in a screening assay to identify/evaluate compounds.For example, a newly synthesized or identified compound (i.e., testcompound) which is labeled can be evaluated for its ability to bind abeta-secretase or a neuron cell (or a neuron cell in the presence of oneor more of beta-amyloid oligomers) by monitoring its concentrationvariation when contacting with the beta-secretase or the neuron cell (orthe neuron cell in the presence of one or more of beta-amyloidoligomers), through tracking the labeling. For another example, a testcompound (labeled) can be evaluated for its ability to reduce binding ofanother compound which is known to bind to beta-secretase or neuron cell(i.e., standard compound). Accordingly, the ability of a test compoundto compete with the standard compound for binding to the beta-secretaseor the neuron cell directly correlates to its binding affinity.Conversely, in some other screening assays, the standard compound islabeled and test compounds are unlabeled. Accordingly, the concentrationof the labeled standard compound is monitored in order to evaluate thecompetition between the standard compound and the test compound, and therelative binding affinity of the test compound is thus ascertained.

Kits

Embodiments of the present invention also includes pharmaceutical kitsuseful, for example, in the treatment or prevention of cognitive declineand/or Alzheimer's disease which include one or more containerscontaining a pharmaceutical composition comprising a therapeuticallyeffective amount of a compound of the invention. Such kits can furtherinclude, if desired, one or more of various conventional pharmaceuticalkit components, such as, for example, containers with one or morepharmaceutically acceptable carriers, additional containers, etc., aswill be readily apparent to those skilled in the art. Instructions,either as inserts or as labels, indicating quantities of the componentsto be administered, guidelines for administration, and/or guidelines formixing the components, can also be included in the kit.

The invention will be described in greater detail by way of specificexamples. The following examples are offered for illustrative purposes,and are not intended to limit the invention in any manner. Those ofskill in the art will readily recognize a variety of non-criticalparameters which can be changed or modified to yield essentially thesame results. Certain compounds of the Examples were found to beinhibit, treat, or abate one or more of amyloid production, amyloidassembly, the activity/effect of Abeta oligomers on neurons (such asneurons in the brain), amyloid aggregation, amyloid oligomer binding,and amyloid deposition according to one or more of the assays providedherein. In some further embodiments, certain compounds of the Exampleswere found to be Inhibit, treat, or abate one or more of theactivity/effect of Abeta oligomers on neurons (such as neurons in thebrain), amyloid aggregation, amyloid (including amyloid oligomer)binding, and amyloid deposition according to one or more of the assaysprovided herein.

In some embodiments, the compound of invention has an IC₅₀ value of lessthan 100 μM, 50 μM, 20 μM, 15 μM, 10 μM, 5 μM, 1 μM, 500 nM, 100 nM, 50nM, or 10 nM with respect to inhibition of one or more of theactivity/effect of Abeta oligomers on neurons (such as neurons in thebrain), amyloid aggregation, amyloid (including amyloid oligomer)binding, and amyloid deposition. In some embodiments, the compound ofinvention has an IC₅₀ value of less than 100 μM, 50 μM, 20 μM, 15 μM, 10μM, 5 μM, 1 μM, 500 nM, 100 nM, 50 nM, or 10 nM with respect toinhibition the activity/effect of Abeta oligomers on neurons (such asneurons in the brain).

In some embodiments, percentage inhibition of the compound of inventionto one or more of the activity/effect of Abeta oligomers on neurons(such as neurons in the brain), amyloid aggregation, amyloid (includingamyloid oligomer) binding, and amyloid deposition was measured at aconcentration of from 10 nM to 10 μM. In some embodiments, thepercentage inhibition measured is about 1% to about 20%, about 20% toabout 50%, about 1% to about 50%, or about 1% to about 80%.

The invention may be appreciated in certain aspects with reference tothe following examples, offered by way of illustration, not by way oflimitation. Materials, reagents and the like to which reference is madein the following examples are obtainable from commercial sources, unlessotherwise noted.

EXAMPLES Materials And Methods Ginger Oil

The light oil extract from ginger root was obtained by supercritical CO₂extraction.

Ginger Oleoresin

The heavy remainder oil was obtained following extraction of ginger rootby supercritical CO₂ extraction.

Example 1 A. Conditioned Extraction of Ginger Oil: Reaction of GingerOil with 4-Chlorobenzylamine Followed by Reduction with SodiumBorohydride in Methanol and by Fractioning Using Column Chromatography

Ginger oil (10 g) was dissolved in toluene (250 mL) and4-chlorobenzylamine (3.4 g) was added. The mixture was maintained underan atmosphere of nitrogen and heated at reflux with removal of water byDean-Stark distillation for 16 hours. At this time the Dean-Stark trapwas removed and the reaction mixture was cooled to 00° C. on an icebath. A solution of sodium borohydride (10 g) in methanol (100 mL) wasadded portion-wise over 30 minutes with vigorous stirring. When theaddition was complete the mixture was heated to reflux for 16 hours. Atthis time the reaction mixture was cooled to room temperature and pouredinto saturated aqueous sodium bicarbonate solution (300 mL). Theresulting mixture was concentrated by rotary evaporation and the aqueousresidue was partitioned between water and chloroform. The chloroformlayer was dried over anhydrous sodium sulfate and then filtered andconcentrated. The products were then fractionated using silica gelcolumn chromatography employing a gradient from 100% chloroform tochloroform:methanol (5:1). The product was detected in the relativelypolar fractions by thin layer chromatography (TLC). Product-containingfractions were combined and concentrated then dried under high vacuumovernight to provide a light brown oil (0.672 g). ¹H NMR (500 MHz,CDCl₃) δ: 7.30-7.24 (m, 4H), 6.81 (d, J=7.8 Hz, 1H), 6.66-6.62 (m, 2H),4.25 (br s, 2H), 3.82 (s, 3H), 3.82 (d, J=13.2 Hz, 1H), 3.72 (d, J=13.2Hz, 1H), 2.73 (m, 1H), 2.66-2.51 (m, 1H), 1.86-1.78 (m, 1H), 1.72-1.63(m, 1H), 1.62-1.51 (m, 1H), 1.17 (d, J=6.3 Hz, 3H). ¹³C NMR (125 MHz,CDCl₃) δ: 146.6, 143.8, 133.9 132.8, 129.9, 129.7, 128.6, 120.8, 114.5,110.9, 55.8, 51.9, 50.2, 38.5, 31.9, 31.6, 29.7, 26.9, 22.6, 19.9.Analytic MS(M+H*): m/z 320.2.

¹H NMR (500 MHz, CD₃OD) δ: 7.10-7.30 (m, 4H), 6.63 (br s, 1H), 6.58 (m,1H), 6.48 (m, 1H), 3.68 (s, 3H), 3.65 (m, 1H), 3.58 (m, 1H), 2.57 (m,1H), 2.50 (m, 1H), 2.35 (m, 1H), 1.73 (m, 1H), 1.49 (m, 1H), 1.04 (d,3H). ¹³C NMR (125 MHz, CD₃OD) δ: 147.5, 145.3, 137.2, 133.3 132.6,129.9, 128.1, 120.4, 114.7, 111.6, 54.9, 51.3, 49.2, 37.7, 31.5, 18.0.

The weight ratio of Ginger oil to 4-chlorobenzylamine used in thereductive amination is about 3:1 (from 2.7:1 to 3.3:1). The chemicalshift measure by ¹H NMR may vary, for example, up to 0.2 ppm. Thechemical shift measure by ¹³H NMR may vary, for example, up to 0.5 ppm.The analytical Mass Spectrum may have an experimental error of +/−0.3.

Purity Determination

The purity of the product was measure by HPLC. The major peak ofretention time of 2.22 minutes indicating greater than about 80%, 85%,90, or 95% of purity. The HPLC conditions used are as follows.

HPLC Conditions:

Mobile Phase A: 13.3 mM ammonium formate/6.7 mM formic acid in water

Mobile Phase B: 6 mM ammonium formate/3 mM formic acid in water/CH₃CN(1/9, v/v)

Column: Synergi Fusion-RP 100A Mercury, 2×20 mm, 2.5 micron

-   -   (Phenomenex Part No 00M-4423-B0_CE)

Gradient Program: RT=2.22 minutes

Time, minute % Phase B Flow rate, ml/min 0 100 0.5 1 100 0.5 2.5 40 0.53.4 40 0.5 3.5 100 0.5 4.5 100 0.5

The purity of the product was also measure by ¹H NMR indicating it to bea single compound of a purity of greater than 90% or 95%.

The structure of Compound Example 1 (or Example Compound 1) Isdetermined to be as follows.

Compound Example 1 4-(3-(4-chlorobenzylamino)butyl)-2-methoxyphenol B.Synthesis by Reductive Amination

Vanillylacetone (5.00 g, 25.7 mmol) was dissolved in toluene (250 mL)and 4-chlorobenzylamine (3.82 g, 27.0 mmol) was added. The mixture wasmaintained under an atmosphere of nitrogen and heated at reflux withremoval of water by Dean-Stark distillation for 16 hours. At this timethe Dean-Stark trap was removed and the reaction mixture was cooled to0° C. on an ice bath. A solution of sodium borohydride (5 g) in methanol(100 mL) was added portion-wise over 30 minutes with vigorous stirring.When the addition was complete the mixture was heated at reflux for 16hours. At this time the reaction mixture was cooled to room temperatureand poured into saturated aqueous sodium bicarbonate solution (300 mL).The resulting mixture was concentrated by rotary evaporation and theaqueous residue was partitioned between water and chloroform. Thechloroform layer was dried over anhydrous sodium sulfate and thenfiltered and concentrated. The product was then purified using silicagel column chromatography employing a mobile phase of 5%ammonia-methanol in chloroform. Product-containing fractions werecombined and concentrated then dried under high vacuum overnight toprovide a light brown oil (6.16 g, 75%). ¹H NMR, ¹³C NMR, and MassSpectrum of the product were substantially the same as those in Example1, A (made by the Conditioned Extraction method).

Example 2 A. Conditioned Extraction of Ginger Oil: Reaction of GingerOil with 4-Trifluoromethylbenzylamine Followed by Reduction with SodiumBorohydride in Methanol and by Fractioning Using Column Chromatography

Ginger oil (10 g) was dissolved in toluene (250 mL) and4-trifluoromethylbenzylamine (3.5 g) was added. The mixture wasmaintained under an atmosphere of nitrogen and heated at reflux withremoval of water by Dean-Stark distillation for 16 hours. At this timethe Dean-Stark trap was removed and the reaction mixture was cooled to0° C. on an ice bath. A solution of sodium borohydride (10 g) inmethanol (100 mL) was added portion-wise over 30 minutes with vigorousstirring. When the addition was complete the mixture was heated toreflux for 16 hours. At this time the reaction mixture was cooled toroom temperature and poured into saturated aqueous sodium bicarbonatesolution (300 mL). The resulting mixture was concentrated by rotaryevaporation and the aqueous residue was partitioned between water andchloroform. The chloroform layer was dried over anhydrous sodium sulfateand then filtered and concentrated. The products were then fractionatedusing silica gel column chromatography employing a gradient from 100%chloroform to chloroform:methanol (5:1). The product was detected in therelatively polar fractions by thin layer chromatography (TLC).Product-containing fractions were combined and concentrated then driedunder high vacuum overnight to provide a light brown oil (0.761 g). ¹HNMR (500 MHz, CDCl₃) δ: 7.57 (d, J=7.8 Hz, 2H), 7.43 (d, J=7.9 Hz, 2H),6.82 (d, J=7.3 Hz, 1H), 6.65 (m, 2H), 5.16-4.42 (br s, 2H), 3.90 (d,J=13.7 Hz, 1H), 3.84 (s, 3H), 3.80 (d, J=13.7 Hz, 1H), 2.76-2.70 (m,1H), 2.67-2.55 (m, 2H), 1.84-1.77 (m, 1H), 1.69-1.63 (m, 1H), 1.17 (d,J=6.3 Hz, 3H). ¹³C NMR (125 MHz, CDCl₃) δ: 146.7, 144.6, 143.9, 134.0,129.1, 128.4, 127.5, 125.4, 125.3, 123.2, 120.8, 114.6, 111.0, 55.7,52.1, 50.6, 38.8, 32.0, 20.1. MS (CI) m/z 353 (M⁺).

The weight ratio of ginger oil to 4-trifluoromethylbenzylamine used inthe reductive amination is about 3:1 (from 2.7:1 to 3.3:1). The chemicalshift measure by ¹H NMR may vary, for example, up to 0.2 ppm. Thechemical shift measure by ¹³H NMR may vary, for example, up to 0.5 ppm.The analytical Mass Spectrum may have an experimental error of +/−0.3.

Purity Determination

The purity of the product was measure by HPLC. The major peak ofretention time of 2.22 minutes indicating greater than about 80%, 85%,90, or 95% of purity. The HPLC conditions used are as follows.

HPLC Conditions:

Mobile Phase A: 13.3 mM ammonium formate/6.7 mM formic acid in waterMobile Phase B: 6 mM ammonium formate/3 mM formic acid in water/CH₃CN(1/9, v/v)

Column: Synergi Fusion-RP 100A Mercury, 2×20 mm, 2.5 micron

-   -   (Phenomenex Part No 00M-4423-B0_CE)

Gradient Program: RT=2.22 minutes

Time, minute % Phase B Flow rate, ml/min 0 100 0.5 1 100 0.5 2.5 40 0.53.4 40 0.5 3.5 100 0.5 4.5 100 0.5

The purity of the product was also measure by ¹H NMR indicating it to bea single compound of a purity of greater than 90% or 95%.

The structure of Compound Example 2 (or Example Compound 2) isdetermined to be as follows.

Compound Example 24-(3-(4-(trifluoromethyl)benzylamino)butyl)-2-methoxyphenol B. Synthesisby Reductive Amination

Vanillylacetone (5.00 g, 25.7 mmol) was dissolved in toluene (250 mL)and 4-trifluoromethylbenzylamine (4.73 g, 27.0 mmol) was added. Themixture was maintained under an atmosphere of nitrogen and heated atreflux with removal of water by Dean-Stark distillation for 16 hours. Atthis time the Dean-Stark trap was removed and the reaction mixture wascooled to 0° C. on an ice bath. A solution of sodium borohydride (5 g)in methanol (100 mL) was added portion-wise over 30 minutes withvigorous stirring. When the addition was complete the mixture was heatedat reflux for 16 hours. At this time the reaction mixture was cooled toroom temperature and poured into saturated aqueous sodium bicarbonatesolution (300 mL). The resulting mixture was concentrated by rotaryevaporation and the aqueous residue was partitioned between water andchloroform. The chloroform layer was dried over anhydrous sodium sulfateand then filtered and concentrated. The product was then purified usingsilica gel column chromatography employing a mobile phase of 5%ammonia-methanol in chloroform. Product-containing fractions werecombined and concentrated then dried under high vacuum overnight toprovide a light brown oil (6.72 g, 74%). ¹H NMR, ¹³C NMR, and MassSpectrum of the product were substantially the same as those in Example2, A (made by the Conditioned Extraction method).

Example AA Exocytosis Assay/MTT Assay

Primary neurons from E18 Sprague-Dawley rat embryos are plated atoptimized concentrations in 384 well plates in NB media (Invitrogen).Neurons are maintained in cultures for 3 weeks, with twice weeklyfeeding of NB media with N₂ supplement (Invitrogen). A test compound isadded to cells, followed by addition of Vehicle or Abeta oligomerpreparations (1.5 μM), and incubated for 1 to 24 hr at 37° C. in 5% CO₂.MTT reagent (3-(4,5-dimethyithizaol-2yl)-2,5diphenyl tetrazoliumbromide) (Roche Molecular Biochemicals) is reconstituted in phosphatebuffered saline to 5 mg/mL. 10 μL of MTT labeling reagent is added toeach well and incubated at 37° C. for 1 h, then imaged.

Each assay plate is formatted so that compounds are tested with andwithout Abeta on each plate. This design eliminates toxic ormetabolically active compounds early on in the screening cascade (at thelevel of the primary screen). Statistical performance of the screeningplate layout are assessed, screening will be initiated if the currentperformance is maintained.

Similar procedures for exocytosis assays/MTT assays can be found in theliterature. See e.g., Liu Y, et. al., Detecting bioactive amyloid betapeptide species in Alzheimer's disease. J Neurochem. 2004 November;91(3):648-56; Liu Y, and Schubert D. “Cytotoxic amyloid peptides inhibitcellular 3-(4.5-dimethylthiazol-2-yl)-2,5-diphenyitetrazolium bromide(MTT) reduction by enhancing MTT formazan exocytosis.” J Neurochem. 1997December; 69(6):2285-93; and Liu Y, and Schubert D. “TreatingAlzheimer's disease by inactivating bioactive amyloid beta peptide”Curr. Alzheimer Res. 2006 April; 3(2):129-35.

Experimental Controls:

Abeta 1-42 oligomers made according to published methods methods [Seee.g. Dahlgren et al., “Oligomeric and fibrillar species of amyloid-betapeptides differentially affect neuronal viability” J Biol Chem. 2002Aug. 30; 277(35):32046-53. Epub 2002 Jun. 10; LeVine H 3rd. “Alzheimer'sbeta-peptide oligomer formation at physiologic concentrations” AnalBiochem. 2004 Dec. 1; 335(1):81-90; Shrestha et. al, “Amyloid betapeptide adversely affects spine number and motility in hippocampalneurons” Mol Cell Neurosci. 2006 November; 33(3):274-82. Epub 2006 Sep.8; Puzzo et al., “Amyloid-beta peptide inhibits activation of the nitricoxide/cGMP/cAMP-responsive element-binding protein pathway duringhippocampal synaptic plasticity” J Neurosci. 2005 Jul. 20;25(29):6887-97; Barghorn et al., “Globular amyloid beta-peptideoligomer—a homogenous and stable neuropathological protein inAlzheimer's disease” J Neurochem. 2005 November; 95(3):834-47. Epub 2005Aug. 31; Johansson et al., Physiochemical characterization of theAlzheimer's disease-related peptides A beta 1-42 Arctic and A beta 1-42wt. FEBS J. 2006 June; 2 73(12):2618-30] as well as brain-derived Abetaoligomers (See e.g. Walsh et al., Naturally secreted oligomers ofamyloid beta protein potently inhibit hippocampal long-term potentiationin vivo. Nature (2002). 416, 535-539; Lesne et al., A specificamyloid-beta protein assembly in the brain impairs memory. Nature. 2006Mar. 16; 440(7082):352-7; Shankar et al, Amyloid-beta protein dimersisolated directly from Alzheimer's brains impair synaptic plasticity andmemory. Nat Med. 2008 August; 14(8):837-42. Epub 2008 Jun. 22)constitute the positive controls. Negative controls includevehicle-treated neurons as well as neurons treated with 28 μMconcentrations of memantine. Memantine produces 50% inhibition ofoligomer effects at this dose. These controls, on each plate, serve asnormalization tools to calibrate assay performance on a plate-by-platebasis.

Primary Neuronal Cultures

Optimal cell density is determined based on cellular response to Abetaoligomers using the exocytosis assay as a readout, andimmunohistochemical analysis of the relative proportion of glia toneurons in the cultures. Cultures are monitored on a weekly basis withimmunohistochemistry and image processing-based quantification tomonitor the percentage of the cultures that are neurons vs. glia (Glialcells). Cultures containing more than 20% glia (positive for GFAP) vs.neurons (staining positively with antibodies directed against MAP2) atthe screening age of 21 days in vitro (21 DIV) are rejected.

Abeta Oligomer Preparations

Human amyloid peptide 1-42 is obtained from California Peptide, withlot-choice contingent upon quality control analysis. Quality controls ofoligomer preparations consist of Westerns to determine oligomer sizeranges and relative concentrations, and the MTT assay to confirmexocytosis acceleration without toxicity. Toxicity is monitored in eachimage-based assay via quantification of nuclear morphology visualizedwith the DNA binding dye DAPI (Invitrogen). Nuclei that are fragmentedare considered to be in late stage apoptosis (Majno and Joris '95).Peptide lots producing unusual peptide size ranges or significanttoxicity at a standard 1.5 uM concentration on neurons are rejected.Plate-based controls—The assay optimization will be complete whenreformatted plates achieve a minimum of statistically significanttwo-fold separation between vehicle and Abeta oligomer-treated neurons(p<0.01, Student's t-test, unequal variance) on a routine basis, with nomore than 10% CV between plates, equivalent to its current performance.

Statistical Software and Analysis:

Data handling and analysis are accomplished by Cellomics VTI imageanalysis software and STORE automated database software. Because of thelow dynamic range and neuronal well-to-well variability after threeweeks in culture, statistical comparisons are made via pairwiseTukey-Kramer analysis to determine the significance of the separationbetween compound+Abeta oligomers from Abeta alone, and between compoundalone from vehicle. These statistics are more akin to what is seen inanimal behavioral testing than the z′ statistic that has been used forthe past two decades in high throughput screening. The ability of matureprimary neurons to more closely approximate the electrophysiologicallymediated signal transduction network of the adult brain justifies thisscreening strategy. Power analysis will be set for a number of replicatescreening wells that will minimize false negatives (e.g N=4) and shiftthe burden of distinguishing false positives from actual hits todose-response confirmation screening. Rank ordering of compounds is doneon the basis of secondary assay mechanism of action and physicochemicalproperties of the compound structures. Certain test compoundssignificantly reverse the effects of Abeta oligomers but not affectneuronal metabolism.

Compound Example 1 was dosed in the MTT assay and was shown to block theAbeta oligomer-induced acceleration of exocytosis with an EC₅₀ of 10 μM,indicating that Compound Example 1 blocks/abate the activity/effect ofAbeta oligomer on neuron cells.

Example BB Binding Assay

Each test compound was added to a plate followed by an addition of oneor more of Abeta 1-42 Oligomers. The plates were fixed with 3.7%paraformaldehyde in phosphate buffered saline (PBS) for 15 min. Theplate was then washed 3× with PBS for 5 min each. The plates wereblocked at room temperature for 1 hour in 5% goat serum and 0.5% TritonX-100 (CAS number: 9002-93-1) in PBS. Primary antibodies (anti-MAP 2polyclonal, Millipore #AB5622 and anti-Beta Amyloid 6E10 monoclonal,Convance #SIG-39300) were diluted 1:1000 in 5% goat serum with PBS.Primary antibodies were incubated either overnight at 4° C. or 1 hour atRT. The plate was then washed 3× with PBS for 5 min each. Secondaryantibodies (Alex Flor 488 polyclonal, Invitrogen # A11008 and Alexa Flor647 monoclonal, Invitrogen #A21235) were diluted 1:1000 in 5% goat serumwith PBS. Secondary antibodies were incubated at RT for 1 hr. The plateswere washed once with PBS. DAPI (4′,6-diamidino-2-phenylindole,Invitrogen) was then applied at 0.03 μg/μL and incubated at RT for 5min, then washed with PBS. Image process was carried out for analysis.

Similar procedures for binding assays can be found in the literature.See e.g., Look G C, et. al. Discovery of ADDL—targeting small moleculedrugs for Alzheimer's disease. Curr Alzheimer Res. 2007 December;4(5):562-7. Review.

Abeta Oligomer Preparations:

Human amyloid peptide 1-42 is obtained from California Peptide, withlot-choice contingent upon quality control analysis. Abeta 1-42oligomers made according to published methods [See e.g. Dahlgren et al.,“Oligomeric and fibrillar species of amyloid-beta peptidesdifferentially affect neuronal viability” J Biol Chem. 2002 Aug. 30;277(35):32046-53. Epub 2002 Jun. 10; LeVine H 3rd. “Alzheimer'sbeta-peptide oligomer formation at physiologic concentrations” AnalBiochem. 2004 Dec. 1; 335(1):81-90; Shrestha et. al, “Amyloid betapeptide adversely affects spine number and motility in hippocampalneurons” Mol Cell Neurosci. 2006 November; 33(3):274-82. Epub 2006 Sep.8; Puzzo et al., “Amyloid-beta peptide inhibits activation of the nitricoxide/cGMP/cAMP-responsive element-binding protein pathway duringhippocampal synaptic plasticity” J Neurosci. 2005 Jul. 20;25(29):6887-97; Barghorn et al., “Globular amyloid beta-peptideoligomer—a homogenous and stable neuropathological protein inAlzheimer's disease” J Neurochem. 2005 November; 95(3):834-47. Epub 2005Aug. 31; Johansson et al., Physiochemical characterization of theAlzheimer's disease-related peptides A beta 1-42 Arctic and A beta 1-42wt. FEBS J. 2006 June; 2 73(12):2618-30] as well as brain-derived Abetaoligomers (See e.g. Walsh et al., Naturally secreted oligomers ofamyloid beta protein potently inhibit hippocampal long-term potentiationin vivo. Nature (2002). 416, 535-539; Lesne et al., A specificamyloid-beta protein assembly in the brain impairs memory. Nature. 2006Mar. 16; 440(7082):352-7; Shankar et al, Amyloid-beta protein dimersisolated directly from Alzheimer's brains impair synaptic plasticity andmemory. Nat Med. 2008 August; 14(8):837-42. Epub 2008 Jun. 22) willserve as positive controls. Quality controls of oligomer preparationsconsist of Westerns to determine oligomer size ranges and relativeconcentrations, and the MTT assay to confirm exocytosis accelerationwithout toxicity. Toxicity is monitored in each image-based assay viaquantification of nuclear morphology visualized with the DNA binding dyeDAPI (Invitrogen). Nuclei that are fragmented are considered to be inlate stage apoptosis (Majno and Joris Apoptosis, oncosis, and necrosis.An overview of cell death. Am J Pathol 1995; 146:3-16). Peptide lotsproducing unusual peptide size ranges or significant toxicity atstandard concentrations on neurons are rejected.

Image Processing

Images were captured and analyzed with the Cellomics VTI automatedmicroscope platform, using the Neuronal Profiling algorithm. Forstatistical analysis, a Tukey-Kramer pair-wise comparison with unequalvariance was used.

Western Blots

Samples containing Abeta 1-42 were diluted (1:5) in non-reducing lanemarker sample buffer (Pierce #1859594). A 30 microliter (μL) sample wasloaded onto an eighteen well precast 4-15% Tris-HCl gel (BIORAD#345-0028). Electrophoresis was performed in a BIO-RAD Criterian precastgel system using Tris-Glycine buffer at 125 volt (V) for 90 minutes. Thegels were blotted onto 0.2 μM nitrocellulose membranes inTris-Glycine/10% methanol buffer at 30V for 120 minutes. The membraneswere boiled for 5 minutes in a PBS solution and blocked over night withTBS/5% milk solution at 4° C. The membrane was probed with 6E10-HRP(Covance #SIG-39345) diluted to 10 μg/mL in TBS/1% milk solution for onehour at room temperature. Membrane was washed three times for 40 minuteseach with a solution of TBS/0.05% tween-20 and developed with ECLreagent (BIO-RAD #162-0112) for 5 minutes. Image acquisition wasperformed on an Alpha Innotech FluorChem Q quantitative imaging systemand analyzed with AlphaView Q software.

PK Studies:

PK studies are performed at CEREP Inc of Redmond Wash., according totheir standard protocols: The plasma samples were processed usingacetonitrile precipitation and analyzed by HPLC-MS or HPLC-MS/MS. Peakareas were recorded, and the concentrations of the test compound in theunknown plasma samples were determined using the respective calibrationcurve. The reportable linear range of the assay was determined, alongwith the lower limit of quantitation (LLQ).

NMR Spectroscopy and Mass Spectrometry:

Active fractions were analyzed by 1H NMR (Varian 500 MHz NMRspectrometer) and purified compounds were characterized using acombination 1D and 2D 1H NMR experiments and 13C NMR experiments.Structure proof was obtained using these NMR techniques in combinationwith low resolution mass spectrometry to determine molecular weight andhigh resolution mass spectrometry (Thermo Finnigan LCQ Ion trap) todetermine composition-of-matter.

Compound Example 1 was shown to partially block binding of the Abetaoligomer ligand to neurons by 24% according to the binding assay (usingimaging processing algorithm).

Example CC Pharmacokinetic Studies

Pharmacokinetic studies were performed according to the followingprotocols: The plasma samples were processed using acetonitrileprecipitation and analyzed by HPLC-MS or HPLC-MS/MS. Peak areas wererecorded, and the concentrations of the test compound in the unknownplasma samples were determined using the respective calibration curve.The reportable linear range of the assay was determined, along with thelower limit of quantitation (LLQ). For example, Compound Example 1 wasdetermined to have a half life of 46 minutes in the plasma of rats wheninjected intravenously at 1 mg/Kg.

Example DD A Primary Neuron-Based Functional Screening Assay to DetectSmall Molecule Abets Oligomer Blockers

Primary rat neurons grown for at least 3 weeks in vitro were chosen asthe basis for this screening assay. These neurons express the fullcomplement of synaptic proteins characteristic of neurons in the maturebrain, and exhibit a complex network of activity-dependent electricalsignaling. Neurons and glia in such cultures have molecular signalingnetworks exhibiting excellent registration with intact brain circuitry,and for this reason have been used for over two decades as a modelsystem for learning and memory (See e.g. Kaech S, Banker G. Culturinghippocampal neurons. Nat Protoc. 2006; 1(5):2406-15. Epub 2007 Jan. 11;See also Craig A M, Graf E R, Linhoff M W. How to build a centralsynapse: clues from cell culture. Trends Neurosci. 2006 Jan. 29(1):8-20.Epub 2005 Dec. 7. Review). More complex systems such as acute ororganotypic brain slices are very useful but not amenable to highthroughput screening. Immortalized or transformed neuronal cell linesare amenable to high throughput screening, but do not replicate theelectrophysiological state-dependent signaling of primary neuronalcultures and are unlikely to adequately model the subtle alterations inthis signaling that are caused by oligomers during the earliestmanifestations of the disease state (See e.g. Görtz P, Flelscher W,Rosenbaum C, Otto F, Siebler M. Neuronal network properties of humanteratocarcinoma cell line-derived neurons. Brain Res. 2004 Aug. 20;1018(1):18-25). For this reason, primary neuronal cultures were chosenbecause of their ability to be used in high throughput screens andfidelity to what occurs in vivo.

Reduced formazan was first visible in intracellular vesicles (FIG. 1A).Example of neurons filled with labeled vesicles following endocytosis ofdye and reduction to an insoluble purple product. (Scale bar=20 micronsin FIG. 1A). Eventual formazan exocytosis was accelerated via Abetaoligomers in mature hippocampal neurons in vitro (FIG. 1B). Examplephotomicrograph of neurons covered with insoluble purple dye that havebeen extruded via exocytosis. The dye precipitated in the aqueousenvironment of the culture and formed needle-shaped crystals on thesurface of the neuron. (FIG. 1B). Endocytosis rate was altered in thepresence of Abeta oligomers. (FIG. 1C) Exocytosis rate was altered inthe presence of Abeta oligomers (FIG. 1D).

Since synaptic and memory deficits, and not widespread cell death,predominate at the earliest stages of Alzheimer's disease, assays thatmeasure these changes can be used to discover small molecule inhibitorsof oligomer activity. The MTT assay can be used as a measure of toxicityin cultures. Yellow tetrazolium salts were endocytosed by cells andreduced to insoluble purple formazan in the endosomal pathway. The levelof purple formazan was a reflection of the number of activelymetabolizing cells in culture, and reduction in the amount of formazanwas taken as a measure of cell death or metabolic toxicity in culture.When observed through a microscope, the purple formazan was firstvisible in intracellular vesicles that fill the cell (Figure. 1A). Overtime, the vesicles were exocytosed and the formazan precipitated asneedle-shaped crystals on the outer surface of the plasma membrane asthe insoluble formazan was exposed to the aqueous media environment(FIG. 1B). Cells respond to sublethal levels of Abeta oligomers byselectively accelerating the exocytosis rate of reduced formazan, whileleaving endocytosis rate unaffected, which can be seen in mature primaryneurons in vitro and quantified these morphological shifts via automatedmicroscopy and image processing. At a given point in time followingtetrazolium salt addition to the culture well, vehicle-treated cells hadthe appearance of those in FIG. 1A, while Abeta oligomer-treated cellshad the appearance of those in FIG. 1B. Under these circumstances, therewas no overall change in the total amount of reduced formazan, simply ashift in its morphology. This assay is sensitive to low levels ofoligomers that do not cause cell death.

Evidence suggests that Abeta oligomer-mediated reduction in neuronalsurface receptor expression mediated by membrane trafficking are thebasis for oligomer inhibition of electrophysiological measures ofsynaptic plasticity (LTP) and thus learning and memory (See Kamenetz F,Tomita T, Hsieh H, Seabrook G, Borchelt D, Iwatsubo T, Sisodia S.Malinow R. APP processing and synaptic function. Neuron. 2003 Mar. 27;37(6):925-37; and Hsieh H, Boehm J, Sato C, Iwatsubo T, Tomita T,Sisodia S, Malinow R. AMPAR removal underlies Abeta-induced synapticdepression and dendritic spine loss. Neuron. 2006 Dec. 7; 52(5):831-43).Measuring membrane trafficking rate changes induced by oligomers viaformazan morphological shifts has been used in cell lines to discoverAbeta oligomer-blocking drugs [Maezawa I, Hong H S, Wu H C, Battina S K,Rana S, Iwamoto T, Radke G A, Pettersson E, Martin G M, Hua D H, Jin LW. A novel tricyclic pyrone compound ameliorates cell death associatedwith intracellular amyloid-beta oligomeric complexes. J Neurochem. 2006July; 98(1):57-67; Liu Y, Schubert D. Cytotoxic amyloid peptides inhibitcellular 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide(MTT) reduction by enhancing MTT formazan exocytosis. J Neurochem. 1997December; 69(6):2285-93; Liu Y, Dargusch R, Banh C, Miller C A, SchubertD. Detecting bioactive amyloid beta peptide species in Alzheimer'sdisease. J Neurochem. 2004 November; 91(3):648-56; Liu Y, Schubert D.Treating Alzheimer's disease by inactivating bioactive amyloid betapeptide. Curr Alzheimer Res. 2006 April; 3(2):129-35; Rana S, Hong H S,Barrigan L, Jin L W, Hua D H. Syntheses of tricyclic pyrones andpyridinones and protection of Abeta-peptide induced MC65 neuronal celldeath. Bioorg Med Chem Lett. 2009 Feb. 1; 19(3):670-4. Epub 2008 Dec.24; and Hong H S, Maezawa I, Budamagunta M, Rana S, Shi A, Vassar R, LiuR, Lam K S, Cheng R H, Hua D H, Voss J C, Jin L W. Candidate anti-Abetafluorene compounds selected from analogs of amyloid imaging agents.Neurobiol Aging. 2008 Nov. 18. (Epub ahead of print)] that lower Abetabrain levels in rodents in vivo [Hong H S, Rana S, Barrigan L, Shi A,Zhang Y, Zhou F, Jin L W, Hua D H. Inhibition of Alzheimer's amyloidtoxicity with a tricyclic pyrone molecule in vitro and in vivo. JNeurochem. 2009 February; 108(4):1097-1108].

The exocytosis assay was adapted for use with mature primary neuronalcultures grown for 3 weeks in vitro. Abeta oligomers caused adose-dependent decrease in the amount of intracellular vesicles (puncta)filled with reduced purple formazan (FIG. 2A, squares; 3 μM dosecorresponds to image in FIG. 2C) as measured via image processing usinga Cellomics VTI automated microscopy system. Increasing the amount ofAbeta oligomers eventually resulted in overt toxicity. Thus, theconcentration of neuroactive Abeta oligomers was much lower than thatcausing cell death. This decrease can be blocked by addingstoichiometric amounts of anti-Abeta monoclonal antibody 6E10 (IgG) tothe cultures prior to oligomer addition (FIG. 2A, circle; the circlecorresponds to image in FIG. 2D; antibody alone [down triangle] has noeffect on the neurons). Several compounds were tested that have beenreported to block the effects of Abeta oligomers, including the sugaralcohol scyllo-inositol (AZD-103), the nAChR antagonist hexamethoniumbromide, and the NMDAR antagonists MK-801 and none were active (Feniliet al., '07, Calabrese et al., '06, LeCor et al., '07).

The assay was optimized for performance in 384-well microtiter plateswith automated liquid handling robotics for compound formatting andassay plate stamping, routinely achieving statistically significanttwo-fold separation between vehicle and Abeta oligomer-treated neurons(Student's t-test, unequal variance). Compounds were added to neuronsfirst, then oligomers were added. When configured in this manner theassay was able to detect compounds that act via disruption of oligomers,inhibition of oligomer binding to neurons, or counteraction of signaltransduction mechanisms of action initiated by oligomer binding.

Compounds were considered active if they significantly blockAbeta-mediated changes in membrane trafficking, but do not significantlyaffect membrane trafficking when dosed on their own. An example is shownin FIG. 2B; Compound Example 2 inhibits oligomer effects on membranetrafficking with an EC50 of 7 μM.

FIG. 2A shows dose-dependent decrease of intracellular formazan-filledvesicles (puncta) caused by Abeta 42 oligomer treatment acceleration ofexocytosis (squares). Oligomer effects were blocked by anti Abeta IgG(circle and up triangle; circle refers to stoich amount of IgG, i.e., 3μM of Aβ and 1.5 μM of IgG; up triangle refers to substoich IgG, i.e., 3μM of AB and 0.5 μM of IgG). IgG itself (down triangle) has no effect.FIG. 2B shows Example Compound 2, which inhibits oligomer effects onmembrane trafficking. FIG. 2C shows representative micrographs of 21 DIVhippocampal neurons in vitro showing oligomer effects membranetrafficking (corresponding to data point 3 μM in FIG. 2A); and FIG. 2Dshows blockade by anti-Abeta antibodies (corresponding to the circle inFIG. 2A). Data were the average of 3 experiments. Scale bar=20 micron inFIG. 2D.

Example EE Fear Conditioning Assay

Compound Example 2 was tested in an animal model of a memory-dependentbehavioral task known as fear conditioning. The study protocol wasdesigned based on published protocols (See e.g. Puzzo D, Privitera L,Leznik E, Fà M, Staniszewski A, Palmeri A, Arancio O. Picomolaramyloid-beta positively modulates synaptic plasticity and memory inhippocampus. J Neurosci. 2008 Dec. 31; 28(53):14537-45.). The formationof contextual memories is dependent upon the integrity of medialtemporal lobe structures such as the hippocampus. In this assay micewere trained to remember that a particular salient context (conditionedstimulus; CS) is associated with an aversive event, in this case a mildfoot shock (the unconditioned stimulus, US). Animals that show goodlearning will express an increase in freezing behavior when placed backinto the same context. This freezing is absent in a novel context.Increased freezing in the context indicates strong hippocampal-dependentmemory formation in animals. Memory tested in Fear Conditioning issensitive to elevations of soluble AD. FIG. 3 shows the results ofadministration of Abeta oligomers (bar labeled with “a”) during trainingresults in memory deficits when animals are tested 24 later, compared tovehicle administration (bar labeled with “b”). Example Compound 2 waseffective at stopping Abeta oligomer mediated effects on membranetrafficking (FIG. 3). When administered to animals prior to Abetaoligomer administration, Example Compound 2 blocked oligomer effects onmemory in a dose-dependent manner. The compound completely blockedoligomer-mediated memory deficits at the 2 pmol dose (FIG. 3, barlabeled with “d”). This behavioral efficacy demonstrates that themembrane trafficking assay is able to predict which compounds will beefficacious in treating the behavioral memory loss caused by oligomers.The fear condition model for memory was performed as described herein.

FIG. 3 shows that Abeta produces significant deficits in memoryformation vs. vehicle (p<0.05) in the contextual fear conditioningmemory task. FIG. 3 shows that the 2 pmol dose of Compound Example2+Abeta (200 nM) completely blocked the effect of Abeta on memory(p<0.05, one way ANOVA, post hoc comparison with Bonferroni correction).No effect of compound alone was observed(data not shown). No adversebehavioral changes were observed at any dose.

Example FF Membrane Trafficking Assay

Abeta assemblies were isolated from patients with Alzheimer's Disease(AD) or from normal patients. The Abeta assemblies were tested for theirability to modulate membrane trafficking. HMW (>100 KDa) Abetsassemblies isolated from AD patients do not affect membrane trafficking(not shown). IMW (10-100 KDa) Abeta assemblies isolated from AD patientssignificantly affect membrane trafficking. (FIG. 4). IMW Abetaassemblies isolated from Age-matched normal individuals do not affectmembrane trafficking (FIG. 4). Compound Example 2 has no effect on Abetaassemblies isolated from Age-matched normal individuals. (FIG. 4).Compound Example 2 significantly blocked the trafficking effects ofAD-brain derived Abeta aseemblies. (FIG. 4).

Although the present invention has been described in considerable detailwith reference to certain preferred versions thereof, other versions arepossible. Therefore, the spirit and scope of the invention should not belimited to the description of the preferred versions described herein.

All features disclosed in the specification, including the abstract anddrawings, and all the steps in any method or process disclosed, may becombined in any combination, except combinations where at least some ofsuch features and/or steps are mutually exclusive. Each featuredisclosed in the specification, including abstract and drawings, can bereplaced by alternative features serving the same, equivalent or similarpurpose, unless expressly stated otherwise. Thus, unless expresslystated otherwise, each feature disclosed is one example only of ageneric series of equivalent or similar features. Various modificationsof the invention, in addition to those described herein, will beapparent to those skilled in the art from the foregoing description.Such modifications are also intended to fall within the scope of theappended claims.

Each reference cited in the present application is herein incorporatedby reference in its entirety.

1. A compound of Formula I:

or pharmaceutically acceptable salt thereof, wherein: R¹ is selectedfrom (A1) and (A2):

R², R³, R⁴, R⁵, and R⁶ are each, independently, selected from H, OH,C₁₋₆ alkoxy, C₁₋₆ haloalkoxy, halo, CN, NO₂, C₁₋₆ alkyl, C₁₋₆ haloalkyl,C₃₋₇ cycloalkyl, NH₂, NH(C₁₋₄ alkyl), NH(C₃₋₇ cycloalkyl), N(C₁₋₄alkyl)₂, NHC(OX)(C₁₋₄ alkyl), SH, S(C₁₋₆ alkyl), C(O)OR^(a), C(O)R^(b),C(O)NR^(c)R^(d), OC(O)R^(b), OC(O)NR^(c)R^(d), NR^(c)R^(d),NR^(c)C(O)R^(b), NR^(c)C(O)OR^(a), NR^(c)S(O)₂R^(b),NR^(c)S(O)₂NR^(c)R^(d), S(O)R^(b), S(O)₂R^(b), and S(O)₂NR^(c)R^(d); R⁷is H, C₁₋₆ alkyl, C₁₋₆ haloalkyl, or C₃₋₇ cycloalkyl; R⁸ is C₁₋₆ alkyl,C₁₋₆ haloalkyl, or C₃₋₇ cycloalkyl; R⁹ is H, C₁₋₆ alkyl, C₁₋₆ haloalkyl,or C₃₋₇ cycloalkyl; R¹⁰ is H; R¹¹ is H; R¹², R¹³, R¹⁴, R¹⁵, and R¹⁶ areeach, independently, selected from H, OH, C₁₋₆ alkoxy, C₁₋₆ haloalkoxy,halo, CN, NO₂, C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₃₋₇ cycloalkyl, NH₂, NH(C₁₋₄alkyl), NH(C₃₋₇ cycloalkyl), N(C₁₋₄ alkyl)₂, NHC(O)(C₁₋₄ alkyl), SH,S(C₁₋₆ alkyl), C(O)OR^(a1), C(O)R^(b1), C(O)NR^(c1) R^(d1), OC(O)R^(b1),OC(O)NR^(c1) R^(d1), NR^(c1) R^(d1), NR^(c1)C(O)R^(b1),NR^(c1)C(O)OR^(a1), NR^(c1)S(O)₂R^(b1), NR^(c1)S(O)₂NR^(c1) R^(d1),S(O)R^(b1), S(O)₂R^(b1), and S(O)₂NR^(c1) R^(d1); each R^(a) isindependently selected from H, C₁₋₆ alkyl, C₁₋₆ haloalkyl, arylalkyl,heteroarylalkyl, cycloalkylalkyl, heterocycloalkylalkyl, C₂₋₆ alkenyl,C₂₋₆ alkynyl, aryl, cycloalkyl, heteroaryl and heterocycloalkyl, whereineach of the C₁₋₆ alkyl, C₁₋₆ haloalkyl, arylalkyl, heteroarylalkyl,cycloalkylalkyl, heterocycloalkylalkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl,aryl, cycloalkyl, heteroaryl and heterocycloalkyl is optionallysubstituted with 1, 2, 3, 4, or 5 substituents independently selectedfrom OH, CN, amino, halo, C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₁₋₆ alkoxy, C₁₋₆haloalkoxy, aryl, arylalkyl, heteroaryl, heteroarylalkyl, cycloalkyl andheterocycloalkyl; each R^(b) is independently selected from H, C₁₋₆alkyl, C₁₋₆ haloalkyl, arylalkyl, heteroarylalkyl, cycloalkylalkyl,heterocycloalkylalkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl, aryl, cycloalkyl,heteroaryl, heterocycloalkyl, arylalkyl, heteroarylalkyl,cycloalkylalkyl and heterocycloalkylalkyl, wherein each of the C₁₋₆alkyl, C₁₋₆ haloalkyl, arylalkyl, heteroarylalkyl, cycloalkylalkyl,heterocycloalkylalkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl, aryl, cycloalkyl,heteroaryl, heterocycloalkyl, arylalkyl, heteroarylalkyl,cycloalkylalkyl and heterocycloalkylalkyl is optionally substituted with1, 2, 3, 4, or 5 substituents independently selected from OH, amino,halo, C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₁₋₄ alkoxy, C₁₋₆ haloalkoxy, aryl,arylalkyl, heteroaryl, heteroarylalkyl, cycloalkyl and heterocycloalkyl;R^(c) and R^(d) are independently selected from H, C₁₋₆ alkyl, C₁₋₆haloalkyl, arylalkyl, heteroarylalkyl, cycloalkylalkyl,heterocycloalkylalkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl, aryl, heteroaryl,cycloalkyl, heterocycloalkyl, arylalkyl, heteroarylalkyl,cycloalkylalkyl and heterocycloalkylalkyl, wherein each of the C₁₋₆alkyl, C₁₋₆ haloalkyl, arylalkyl, heteroarylalkyl, cycloalkylalkyl,heterocycloalkylalkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl, aryl, heteroaryl,cycloalkyl, heterocycloalkyl, arylalkyl, heteroarylalkyl,cycloalkylalkyl and heterocycloalkylalkyl is optionally substituted with1, 2, 3, 4, or 5 substituents independently selected from OH, amino,halo, C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₁₋₆ alkoxy, C₁₋₆ haloalkoxy, aryl,arylalkyl, heteroaryl, heteroarylalkyl, cycloalkyl and heterocycloalkyl;or R^(c) and R^(d) together with the N atom to which they are attachedform a 4-, 5-, 6- or 7-membered heterocycloalkyl group that isoptionally substituted with 1, 2, 3, 4, or 5 substituents independentlyselected from OH, amino, halo, C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₁₋₆ alkoxy,C₁₋₆ haloalkoxy, aryl, arylalkyl, heteroaryl, heteroarylalkyl,cycloalkyl, and heterocycloalkyl; each R^(a1) is independently selectedfrom H, C₁₋₆ alkyl, C₁₋₆ haloalkyl, arylalkyl, heteroarylalkyl,cycloalkylalkyl, heterocycloalkylalkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl,aryl, cycloalkyl, heteroaryl and heterocycloalkyl, wherein each of theC₁₋₆ alkyl, C₁₋₆ haloalkyl, arylalkyl, heteroarylalkyl, cycloalkylalkyl,heterocycloalkylalkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl, aryl, cycloalkyl,heteroaryl and heterocycloalkyl is optionally substituted with 1, 2, 3,4, or 5 substituents independently selected from OH, CN, amino, halo,C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₁₋₆ alkoxy, C₁₋₆ haloalkoxy, aryl,arylalkyl, heteroaryl, heteroarylalkyl, cycloalkyl, andheterocycloalkyl; each R^(b1) is independently selected from H, C₁₋₆alkyl, C₁₋₆ haloalkyl, arylalkyl, heteroarylalkyl, cycloalkylalkyl,heterocycloalkylalkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl, aryl, cycloalkyl,heteroaryl, heterocycloalkyl, arylalkyl, heteroarylalkyl,cycloalkylalkyl and heterocycloalkylalkyl, wherein each of the C₁₋₆alkyl, C₁₋₆ haloalkyl, arylalkyl, heteroarylalkyl, cycloalkylalkyl,heterocycloalkylalkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl, aryl, cycloalkyl,heteroaryl, heterocycloalkyl, arylalkyl, heteroarylalkyl,cycloalkylalkyl and heterocycloalkylalkyl is optionally substituted with1, 2, 3, 4, or 5 substituents independently selected from OH, amino,halo, C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₁₋₆ alkoxy, C₁₋₆haloalkoxy, aryl,arylalkyl, heteroaryl, heteroarylalkyl, cycloalkyl, andheterocycloalkyl; R^(c1) and R^(d2) are independently selected from H,C₁₋₁₀ alkyl, C₁₋₆ haloalkyl, arylalkyl, heteroarylalkyl,cycloalkylalkyl, heterocycloalkylalkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl,aryl, heteroaryl, cycloalkyl, heterocycloalkyl, arylalkyl,heteroarylalkyl, cycloalkylalkyl and heterocycloalkylalkyl, wherein eachof the C₁₋₁₀ alkyl, C₁₋₆ haloalkyl, arylalkyl, heteroarylalkyl,cycloalkylalkyl, heterocycloalkylalkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl,aryl, heteroaryl, cycloalkyl, heterocycloalkyl, arylalkyl,heteroarylalkyl, cycloalkylalkyl and heterocycloalkylalkyl is optionallysubstituted with 1, 2, 3, 4, or 5 substituents independently selectedfrom OH, amino, halo, C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₁₋₆ alkoxy, d-₆haloalkoxy, aryl, arylalkyl, heteroaryl, heteroarylalkyl, cycloalkyl,and heterocycloalkyl; or R^(c1) and R^(d1) together with the N atom towhich they are attached form a 4-, 5-, 6- or 7-membered heterocycloalkylgroup that is optionally substituted with 1, 2, 3, 4, or 5 substituentsindependently selected from OH, amino, halo, C₁₋₆ alkyl, C₁₋₆ haloalkyl,C₁₋₆ alkoxy, C₁₋₆ haloalkoxy, aryl, arylalkyl, heteroaryl,heteroarylalkyl, cycloalkyl, and heterocycloalkyl; and m is 0, 1, or 2,with the proviso that (a) when R¹ is a moiety of (A1), then two of R²,R³, R⁴, R⁵, and R⁶ are independently selected from OH, C₁₋₆ alkoxy, andC₁₋₆ haloalkoxy; and (b) when R¹ is a moiety of (A1), then at least oneof R¹², R¹³, R¹⁴, R¹⁵, and R¹⁶ is other than H.
 2. The compound of claim1 or pharmaceutically acceptable salt thereof, wherein two of R², R³,R⁴, R⁵, and R⁶ are independently selected from OH, C₁₋₆ alkoxy, and C₁₋₆haloalkoxy.
 3. The compound of claim 1 or pharmaceutically acceptablesalt thereof, wherein R², R³, R⁴, R⁵, and R⁶ are each, independently,selected from H, OH, C₁₋₆ alkoxy, C₁₋₆ haloalkoxy, halo, CN, NO₂, C₁₋₆alkyl, C₁₋₆ haloalkyl, C₃₋₇ cycloalkyl, NH₂, NH(C₁₋₄ alkyl), NH(C₃₋₇cycloalkyl), N(C₁₋₄ alkyl)₂, NHC(OX C₁₋₆ alkyl), SH, S(C₁₋₆ alkyl),C(O)OH, C(O)O(C₁₋₄ alkyl), C(O)(C₁₋₄ alkyl), and C(O)NH(C₁₋₄ alkyl). 4.The compound of claim 1 or pharmaceutically acceptable salt thereof,wherein: one of R², R³, R⁴, R⁵, and R⁶ is OH; and one of R², R³, R⁴, R⁵,and R⁶ is OH, C₁₋₆ alkoxy, or C₁₋₆haloalkoxy.
 5. The compound of claim 1or pharmaceutically acceptable salt thereof, wherein: one of R², R³, R⁴,R⁵, and R⁶ is OH; and one of R², R³, R⁴, R⁵, and R⁶ is C₁₋₃ alkoxy orC₁₋₃ haloalkoxy.
 6. The compound of claim 1 or pharmaceuticallyacceptable salt thereof, wherein: one of R², R³, R⁴, R⁵, and R⁶ is OH;and one of R², R³, R⁴, R⁵, and R⁶ is methoxy or trihalomethoxy.
 7. Thecompound of claim 1 or pharmaceutically acceptable salt thereof,wherein: one of R², R³, R⁴, R⁵, and R⁶ is OH; and one of R², R³, R⁴, R⁵,and R⁶ is methoxy.
 8. The compound of claim 1 or pharmaceuticallyacceptable salt thereof, wherein: R⁴ is OH; and R⁵ is methoxy.
 9. Thecompound of claim 1 or pharmaceutically acceptable salt thereof,wherein: R⁴ is OH; R⁵ is methoxy; and R², R³, and R⁶ are each H.
 10. Thecompound of claim 1 or pharmaceutically acceptable salt thereof, whereinR⁷ is H or C₁₋₆ alkyl.
 11. The compound of claim 1 or pharmaceuticallyacceptable salt thereof, wherein R⁷ is H or C₁₋₃ alkyl.
 12. The compoundof claim 1 or pharmaceutically acceptable salt thereof, wherein R⁷ isC₁₋₃ alkyl.
 13. The compound of claim 1 or pharmaceutically acceptablesalt thereof, wherein R⁷ is H.
 14. The compound of claim 1 orpharmaceutically acceptable salt thereof, wherein R⁸ is C₁₋₆ alkyl. 15.The compound of claim 1 or pharmaceutically acceptable salt thereof,wherein R⁸ is C₁₋₃ alkyl.
 16. The compound of claim 1 orpharmaceutically acceptable salt thereof, wherein R⁸ is methyl.
 17. Thecompound of claim 1 or pharmaceutically acceptable salt thereof, whereinR⁹ is H or C₁₋₆ alkyl.
 18. The compound of claim 1 or pharmaceuticallyacceptable salt thereof, wherein R⁹ is H or C₁₋₆ alkyl.
 19. The compoundof claim 1 or pharmaceutically acceptable salt thereof, wherein R⁹ is H.20. The compound of claim 1 or pharmaceutically acceptable salt thereof,wherein R⁹ is C₁₋₃ alkyl.
 21. The compound of claim 1 orpharmaceutically acceptable salt thereof, wherein R¹⁰ is H or C₁₋₆alkyl.
 22. (canceled)
 23. The compound of claim 1 or pharmaceuticallyacceptable salt thereof, wherein R¹⁰ is H.
 24. (canceled)
 25. (canceled)26. The compound of claim 1 or pharmaceutically acceptable salt thereof,wherein R¹¹ is H.
 27. The compound of claim 1 or pharmaceuticallyacceptable salt thereof, wherein at least one of R², R³, R⁴, R¹⁵, andR¹⁶ is other than H.
 28. The compound of claim 1 or pharmaceuticallyacceptable salt thereof, wherein R¹², R¹³, R¹⁴, R¹⁵, and R¹⁶ are each,independently, selected from H, OH, C₁₋₆ alkoxy, C₁₋₆ haloalkoxy, halo,CN, NO₂, C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₃₋₇ cycloalkyl, NH₂, NH(C₁₋₄alkyl), NH(C₃₋₇ cycloalkyl), N(C₁₋₄ alkyl)₂, NHC(O)(C₁₋₆ alkyl), SH,S(C₁₋₆ alkyl), C(O)OH, C(O)O(C₁₋₆ alkyl), C(O)(C₁₋₄ alkyl), andC(O)NH(C₁₋₄ alkyl).
 29. The compound of claim 1 or pharmaceuticallyacceptable salt thereof, wherein R¹², R¹³, R¹⁴, R¹⁵, and R¹⁶ are each,independently, selected from H, halo, CN, NO₂, C₁₋₆ alkyl, C₁₋₆haloalkyl, C₃₋₇ cycloalkyl, C(O)O(C₁₋₄ alkyl), C(O)(C₁₋₄ alkyl), andC(O)NH(C₁₋₄ alkyl).
 30. The compound of claim 1 or pharmaceuticallyacceptable salt thereof, wherein at least one of R¹², R¹³, R¹⁴, R¹⁵, andR¹⁶ is selected from halo, CN, NO₂, C₁₋₆ haloalkyl, C(O)O(C₁₋₆ alkyl),C(O)(C₁₋₄ alkyl), and C(O)NH(C₁₋₄ alkyl).
 31. The compound of claim 1 orpharmaceutically acceptable salt thereof, wherein at least one of R¹²,R¹³, R¹⁴, R¹⁵, and R¹⁶ is selected from halo and C₁₋₆ haloalkyl.
 32. Thecompound of claim 1 or pharmaceutically acceptable salt thereof, whereinR¹⁴ is halo.
 33. The compound of claim 1 or pharmaceutically acceptablesalt thereof, wherein R¹⁴ is C₁₋₆ haloalkyl.
 34. The compound of claim 1or pharmaceutically acceptable salt thereof, wherein R¹⁴ and R¹⁵ areindependently halo.
 35. (canceled)
 36. The compound of claim 35 orpharmaceutically acceptable salt thereof, wherein the compound is acompound of Formula IIa:


37. The compound of claim 35 or pharmaceutically acceptable saltthereof, wherein the compound is a compound of Formula IIb:


38. The compound of claim 1 or pharmaceutically acceptable salt thereof,wherein the compound is a compound of Formula III:


39. The compound of claim 38 or pharmaceutically acceptable saltthereof, wherein m is
 1. 40. The compound of claim 38 orpharmaceutically acceptable salt thereof, wherein m is
 0. 41. (canceled)42. (canceled)
 43. A pharmaceutical composition comprising a compound ofclaim 1 or pharmaceutically acceptable salt thereof, and apharmaceutically acceptable carrier.
 44. (canceled)
 45. (canceled) 46.(canceled)
 47. (canceled)
 48. (canceled)
 49. (canceled)
 50. (canceled)51. (canceled)
 52. (canceled)
 53. A method of inhibiting, treating,and/or abatement of cognitive decline and/or inhibiting Alzheimer'sdisease in a patient comprising administrating to the patient a compoundof claim 1 or a pharmaceutically acceptable salt thereof.
 54. The methodof claim 53 wherein the method comprises inhibiting, treating, orabatement of one or more symptoms of cognitive decline selected from thegroup consisting of memory loss, confusion, impaired judgment,personality changes, disorientation, and loss of language skills. 55.The method of claim 53 wherein the method comprises one or more of: (i)restoration of long term potentiation; and/or (ii) inhibiting, treating,and/or abatement of neurodegeneration; and/or (iii) inhibiting,treating, and/or abatement of general amyloidosis; and/or (iv)inhibiting, treating, and/or abatement of one or more of amyloidproduction, amyloid assembly, amyloid aggregation, amyloid oligomerbinding, and amyloid deposition; and/or (v) inhibiting, treating, and/orabatement of the activity/effect of one or more of Abeta oligomers on aneuron cell.
 56. The method of claim 53 wherein the method comprisesinhibiting, treating, and/or abatement of one or more of amyloidproduction, amyloid assembly, the activity/effect of one or more ofAbeta oligomers on a neuron cell, amyloid aggregation, amyloid binding,and amyloid deposition.
 57. The method of claim 53 wherein the methodcomprises inhibiting, treating, and/or abatement of one or more of theactivity/effect of one or more of Abeta oligomers on a neuron cell,amyloid aggregation, amyloid binding, and amyloid deposition.
 58. Amethod of inhibiting, treating, and/or abatement of cognitive declineand/or inhibiting Alzheimer's disease in a patient comprisingadministrating to the patient a compound of claim 1 or apharmaceutically acceptable salt thereof.
 59. The method of claim 58wherein the method comprises inhibiting, treating, or abatement of oneor more symptoms of cognitive decline selected from the group consistingof memory loss, confusion, impaired judgment, personality changes,disorientation, and loss of language skills.
 60. The method of claim 58wherein the method one or more of: (i) restoration of long termpotentiation; and/or (ii) inhibiting, treating, and/or abatement ofneurodegeneration; and/or (iii) inhibiting, treating, and/or abatementof general amyloidosis; and/or (iv) inhibiting, treating, and/orabatement of one or more of amyloid production, amyloid assembly,amyloid aggregation, amyloid oligomer binding, and amyloid deposition;and/or (v) inhibiting, treating, and/or abatement of the activity/effectof one or more of Abeta oligomers on a neuron cell.
 61. The method ofclaim 58 wherein the method comprises inhibiting, treating, and/orabatement of one or more of amyloid production, amyloid assembly, theactivity/effect of one or more of Abeta oligomers on a neuron cell,amyloid aggregation, amyloid binding, and amyloid deposition.
 62. Themethod of claim 58 wherein the method comprises inhibiting, treating,and/or abatement of one or more of the activity/effect of one or more ofAbeta oligomers on a neuron cell, amyloid aggregation, amyloid binding,and amyloid deposition.